IntAct European Bioinformatics Institute IntAct intact http://www.ebi.ac.uk/ http://www.ebi.ac.uk/intact/query/${ac} EBI-[0-9]+|IA:[0-9]+ INTerAction database (IntAct) provides an open source database and toolkit for the storage, presentation and analysis of molecular interactions. http://www.ebi.ac.uk/intact European Bioinformatics Institute; Wellcome Trust Genome Campus; Hinxton, Cambridge; CB10 1SD; United Kingdom http://www.ebi.ac.uk/intact/ A DNA repair complex functions as an oct4/sox2 coactivator in embryonic stem cells. A DNA repair complex functions as an oct4/sox2 coactivator in embryonic stem cells. Cell (0092-8674) 2011 imex curation Fong Y.W., Inouye C., Yamaguchi T., Cattoglio C., Grubisic I., Tjian R. Nuclear extracts were prepared from NT2 cells, precipitated with ammonium sulfate and resuspended. Soluble fraction was applied to a P11-phosphocellulose column, washed and eluted at 1.0 M KCl (P1M). This P1M fraction was adjusted to 0.05% NP-40 and 10 mM imidazole, applied to a Ni-NTA resin The final Ni-FT fraction was dialyzed and applied to a Poros 20 HQ column and developed with a linear gradient from 0.52 M to 1.0 M KCl. Transcriptionally active fractions were pooled, dialyzed t applied to a Poros 20 HS column, and subjected to a linear gradient from 0.42 M to 1.0 M KCl. Active HS fractions were pooled and applied directly to hydroxyapatite (HAP) type II ceramic resin, and subjected to a linear gradient from 0.19 M to 0.5 M KPi. Active HAP fractions were pooled and separated on a Superose 6 XK 16/70 gel filtration column . Active Superose 6 fractions with an apparent molecular mass of 500-700kDa were pooled and supplemented with 0.25 mg/ml insulin. Pooled fractions were applied to a Poros 20 HE column, and developed with a linear gradient from 0.36 M to 0.86 M KCl. Active HE fractions were supplemented with 0.3 mg/ml insulin, dialyzed and applied to a Mono S PC 1.6/5 column and developed with a linear gradient from 0.2 M to 0.7 M KCl. jmlim@berkeley.edu 21-Dec-2011: Contacted by JYOTI. Only protein-protein interactions human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Fong Y.W., Inouye C., Yamaguchi T., Cattoglio C., Grubisic I., Tjian R. Nuclear extracts were prepared from NT2 cells, precipitated with ammonium sulfate and resuspended. Soluble fraction was applied to a P11-phosphocellulose column, washed and eluted at 1.0 M KCl (P1M). This P1M fraction was adjusted to 0.05% NP-40 and 10 mM imidazole, applied to a Ni-NTA resin The final Ni-FT fraction was dialyzed and applied to a Poros 20 HQ column and developed with a linear gradient from 0.52 M to 1.0 M KCl. Transcriptionally active fractions were pooled, dialyzed t applied to a Poros 20 HS column, and subjected to a linear gradient from 0.42 M to 1.0 M KCl. Active HS fractions were pooled and applied directly to hydroxyapatite (HAP) type II ceramic resin, and subjected to a linear gradient from 0.19 M to 0.5 M KPi. Active HAP fractions were pooled and separated on a Superose 6 XK 16/70 gel filtration column . Active Superose 6 fractions with an apparent molecular mass of 500-700kDa were pooled and supplemented with 0.25 mg/ml insulin. Pooled fractions were applied to a Poros 20 HE column, and developed with a linear gradient from 0.36 M to 0.86 M KCl. Active HE fractions were supplemented with 0.3 mg/ml insulin, dialyzed and applied to a Mono S PC 1.6/5 column and developed with a linear gradient from 0.2 M to 0.7 M KCl. jmlim@berkeley.edu Accepted 2011-NOV-03 by MGT 21-Dec-2011: Contacted by JYOTI. Cell (0092-8674) 2011 Only protein-protein interactions imex curation A DNA repair complex functions as an oct4/sox2 coactivator in embryonic stem cells. A DNA repair complex functions as an oct4/sox2 coactivator in embryonic stem cells. Cell (0092-8674) 2011 imex curation Fong Y.W., Inouye C., Yamaguchi T., Cattoglio C., Grubisic I., Tjian R. Nuclear extracts were prepared from NT2 cells, precipitated with ammonium sulfate and resuspended. Soluble fraction was applied to a P11-phosphocellulose column, washed and eluted at 1.0 M KCl (P1M). This P1M fraction was adjusted to 0.05% NP-40 and 10 mM imidazole, applied to a Ni-NTA resin The final Ni-FT fraction was dialyzed and applied to a Poros 20 HQ column and developed with a linear gradient from 0.52 M to 1.0 M KCl. Transcriptionally active fractions were pooled, dialyzed t applied to a Poros 20 HS column, and subjected to a linear gradient from 0.42 M to 1.0 M KCl. Active HS fractions were pooled and applied directly to hydroxyapatite (HAP) type II ceramic resin, and subjected to a linear gradient from 0.19 M to 0.5 M KPi. Active HAP fractions were pooled and separated on a Superose 6 XK 16/70 gel filtration column . Active Superose 6 fractions with an apparent molecular mass of 500-700kDa were pooled and supplemented with 0.25 mg/ml insulin. Pooled fractions were applied to a Poros 20 HE column, and developed with a linear gradient from 0.36 M to 0.86 M KCl. Active HE fractions were supplemented with 0.3 mg/ml insulin, dialyzed and applied to a Mono S PC 1.6/5 column and developed with a linear gradient from 0.2 M to 0.7 M KCl. jmlim@berkeley.edu 21-Dec-2011: Contacted by JYOTI. Only protein-protein interactions spofr-sf_9 Spodoptera frugiperda insect cells Fall armyworm sf_9 insect cells Derived from pupal ovarian tissue of the fall armyworm Cloned from the parent cell line, IPLB-SF-21-AE (SF-21) derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae cells are susceptible to Baculovirus infection affinity chrom affinity chromatography technology Affinity purification Affinity purification affinity chrom weight by comassie molecular weight estimation by coomasie staining weight by comassie Fong Y.W., Inouye C., Yamaguchi T., Cattoglio C., Grubisic I., Tjian R. Only coomassie staining but I think its OK as they run the purified proteins, which have been ID'd by mass spec and western on the same gel, and its a very clean purification. Nuclear extracts were prepared from NT2 cells, precipitated with ammonium sulfate and resuspended. Soluble fraction was applied to a P11-phosphocellulose column, washed and eluted at 1.0 M KCl (P1M). This P1M fraction was adjusted to 0.05% NP-40 and 10 mM imidazole, applied to a Ni-NTA resin The final Ni-FT fraction was dialyzed and applied to a Poros 20 HQ column and developed with a linear gradient from 0.52 M to 1.0 M KCl. Transcriptionally active fractions were pooled, dialyzed t applied to a Poros 20 HS column, and subjected to a linear gradient from 0.42 M to 1.0 M KCl. Active HS fractions were pooled and applied directly to hydroxyapatite (HAP) type II ceramic resin, and subjected to a linear gradient from 0.19 M to 0.5 M KPi. Active HAP fractions were pooled and separated on a Superose 6 XK 16/70 gel filtration column . Active Superose 6 fractions with an apparent molecular mass of 500-700kDa were pooled and supplemented with 0.25 mg/ml insulin. Pooled fractions were applied to a Poros 20 HE column, and developed with a linear gradient from 0.36 M to 0.86 M KCl. Active HE fractions were supplemented with 0.3 mg/ml insulin, dialyzed and applied to a Mono S PC 1.6/5 column and developed with a linear gradient from 0.2 M to 0.7 M KCl. jmlim@berkeley.edu Accepted 2011-NOV-03 by MGT 21-Dec-2011: Contacted by JYOTI. Cell (0092-8674) 2011 Only protein-protein interactions imex curation A DNA repair complex functions as an oct4/sox2 coactivator in embryonic stem cells. A DNA repair complex functions as an oct4/sox2 coactivator in embryonic stem cells. Cell (0092-8674) 2011 imex curation Fong Y.W., Inouye C., Yamaguchi T., Cattoglio C., Grubisic I., Tjian R. Nuclear extracts were prepared from NT2 cells, precipitated with ammonium sulfate and resuspended. Soluble fraction was applied to a P11-phosphocellulose column, washed and eluted at 1.0 M KCl (P1M). This P1M fraction was adjusted to 0.05% NP-40 and 10 mM imidazole, applied to a Ni-NTA resin The final Ni-FT fraction was dialyzed and applied to a Poros 20 HQ column and developed with a linear gradient from 0.52 M to 1.0 M KCl. Transcriptionally active fractions were pooled, dialyzed t applied to a Poros 20 HS column, and subjected to a linear gradient from 0.42 M to 1.0 M KCl. Active HS fractions were pooled and applied directly to hydroxyapatite (HAP) type II ceramic resin, and subjected to a linear gradient from 0.19 M to 0.5 M KPi. Active HAP fractions were pooled and separated on a Superose 6 XK 16/70 gel filtration column . Active Superose 6 fractions with an apparent molecular mass of 500-700kDa were pooled and supplemented with 0.25 mg/ml insulin. Pooled fractions were applied to a Poros 20 HE column, and developed with a linear gradient from 0.36 M to 0.86 M KCl. Active HE fractions were supplemented with 0.3 mg/ml insulin, dialyzed and applied to a Mono S PC 1.6/5 column and developed with a linear gradient from 0.2 M to 0.7 M KCl. jmlim@berkeley.edu 21-Dec-2011: Contacted by JYOTI. Only protein-protein interactions human-nt2 Homo sapiens neuronal precursor cells nt2 NT2 cells-human neuronal precursor cell line Source of these cells was Stratagene. They were derived from a lung metastasis of a 22 year old Caucasian male with a testicular germ cell tumor. Transplantation into nude mice resulted in a tumor that contained embryonal carcinoma cells. The Ntera-2 (NT2) line was established from this tumor.(taken from the Stratagene info on the web). chromatography chromatography technology column chromatography chromatography western blot western blot Immuno blot Immuno blot Fong Y.W., Inouye C., Yamaguchi T., Cattoglio C., Grubisic I., Tjian R. Nuclear extracts were prepared from NT2 cells, precipitated with ammonium sulfate and resuspended. Soluble fraction was applied to a P11-phosphocellulose column, washed and eluted at 1.0 M KCl (P1M). This P1M fraction was adjusted to 0.05% NP-40 and 10 mM imidazole, applied to a Ni-NTA resin The final Ni-FT fraction was dialyzed and applied to a Poros 20 HQ column and developed with a linear gradient from 0.52 M to 1.0 M KCl. Transcriptionally active fractions were pooled, dialyzed t applied to a Poros 20 HS column, and subjected to a linear gradient from 0.42 M to 1.0 M KCl. Active HS fractions were pooled and applied directly to hydroxyapatite (HAP) type II ceramic resin, and subjected to a linear gradient from 0.19 M to 0.5 M KPi. Active HAP fractions were pooled and separated on a Superose 6 XK 16/70 gel filtration column . Active Superose 6 fractions with an apparent molecular mass of 500-700kDa were pooled and supplemented with 0.25 mg/ml insulin. Pooled fractions were applied to a Poros 20 HE column, and developed with a linear gradient from 0.36 M to 0.86 M KCl. Active HE fractions were supplemented with 0.3 mg/ml insulin, dialyzed and applied to a Mono S PC 1.6/5 column and developed with a linear gradient from 0.2 M to 0.7 M KCl. jmlim@berkeley.edu Accepted 2011-NOV-03 by MGT 21-Dec-2011: Contacted by JYOTI. Cell (0092-8674) 2011 Only protein-protein interactions imex curation A DNA repair complex functions as an oct4/sox2 coactivator in embryonic stem cells. A DNA repair complex functions as an oct4/sox2 coactivator in embryonic stem cells. Cell (0092-8674) 2011 imex curation Fong Y.W., Inouye C., Yamaguchi T., Cattoglio C., Grubisic I., Tjian R. Nuclear extracts were prepared from NT2 cells, precipitated with ammonium sulfate and resuspended. Soluble fraction was applied to a P11-phosphocellulose column, washed and eluted at 1.0 M KCl (P1M). This P1M fraction was adjusted to 0.05% NP-40 and 10 mM imidazole, applied to a Ni-NTA resin The final Ni-FT fraction was dialyzed and applied to a Poros 20 HQ column and developed with a linear gradient from 0.52 M to 1.0 M KCl. Transcriptionally active fractions were pooled, dialyzed t applied to a Poros 20 HS column, and subjected to a linear gradient from 0.42 M to 1.0 M KCl. Active HS fractions were pooled and applied directly to hydroxyapatite (HAP) type II ceramic resin, and subjected to a linear gradient from 0.19 M to 0.5 M KPi. Active HAP fractions were pooled and separated on a Superose 6 XK 16/70 gel filtration column . Active Superose 6 fractions with an apparent molecular mass of 500-700kDa were pooled and supplemented with 0.25 mg/ml insulin. Pooled fractions were applied to a Poros 20 HE column, and developed with a linear gradient from 0.36 M to 0.86 M KCl. Active HE fractions were supplemented with 0.3 mg/ml insulin, dialyzed and applied to a Mono S PC 1.6/5 column and developed with a linear gradient from 0.2 M to 0.7 M KCl. jmlim@berkeley.edu 21-Dec-2011: Contacted by JYOTI. Only protein-protein interactions spofr-sf_9 Spodoptera frugiperda insect cells Fall armyworm sf_9 insect cells Derived from pupal ovarian tissue of the fall armyworm Cloned from the parent cell line, IPLB-SF-21-AE (SF-21) derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae cells are susceptible to Baculovirus infection pull down pull down weight by comassie molecular weight estimation by coomasie staining weight by comassie Fong Y.W., Inouye C., Yamaguchi T., Cattoglio C., Grubisic I., Tjian R. Only coomassie staining but I think its OK as they run the purified proteins, which have been ID'd by mass spec and western on the same gel, and its a very clean purification. Nuclear extracts were prepared from NT2 cells, precipitated with ammonium sulfate and resuspended. Soluble fraction was applied to a P11-phosphocellulose column, washed and eluted at 1.0 M KCl (P1M). This P1M fraction was adjusted to 0.05% NP-40 and 10 mM imidazole, applied to a Ni-NTA resin The final Ni-FT fraction was dialyzed and applied to a Poros 20 HQ column and developed with a linear gradient from 0.52 M to 1.0 M KCl. Transcriptionally active fractions were pooled, dialyzed t applied to a Poros 20 HS column, and subjected to a linear gradient from 0.42 M to 1.0 M KCl. Active HS fractions were pooled and applied directly to hydroxyapatite (HAP) type II ceramic resin, and subjected to a linear gradient from 0.19 M to 0.5 M KPi. Active HAP fractions were pooled and separated on a Superose 6 XK 16/70 gel filtration column . Active Superose 6 fractions with an apparent molecular mass of 500-700kDa were pooled and supplemented with 0.25 mg/ml insulin. Pooled fractions were applied to a Poros 20 HE column, and developed with a linear gradient from 0.36 M to 0.86 M KCl. Active HE fractions were supplemented with 0.3 mg/ml insulin, dialyzed and applied to a Mono S PC 1.6/5 column and developed with a linear gradient from 0.2 M to 0.7 M KCl. jmlim@berkeley.edu Accepted 2011-NOV-03 by MGT 21-Dec-2011: Contacted by JYOTI. Cell (0092-8674) 2011 Only protein-protein interactions imex curation A DNA repair complex functions as an oct4/sox2 coactivator in embryonic stem cells. A DNA repair complex functions as an oct4/sox2 coactivator in embryonic stem cells. Cell (0092-8674) 2011 imex curation Fong Y.W., Inouye C., Yamaguchi T., Cattoglio C., Grubisic I., Tjian R. Nuclear extracts were prepared from NT2 cells, precipitated with ammonium sulfate and resuspended. Soluble fraction was applied to a P11-phosphocellulose column, washed and eluted at 1.0 M KCl (P1M). This P1M fraction was adjusted to 0.05% NP-40 and 10 mM imidazole, applied to a Ni-NTA resin The final Ni-FT fraction was dialyzed and applied to a Poros 20 HQ column and developed with a linear gradient from 0.52 M to 1.0 M KCl. Transcriptionally active fractions were pooled, dialyzed t applied to a Poros 20 HS column, and subjected to a linear gradient from 0.42 M to 1.0 M KCl. Active HS fractions were pooled and applied directly to hydroxyapatite (HAP) type II ceramic resin, and subjected to a linear gradient from 0.19 M to 0.5 M KPi. Active HAP fractions were pooled and separated on a Superose 6 XK 16/70 gel filtration column . Active Superose 6 fractions with an apparent molecular mass of 500-700kDa were pooled and supplemented with 0.25 mg/ml insulin. Pooled fractions were applied to a Poros 20 HE column, and developed with a linear gradient from 0.36 M to 0.86 M KCl. Active HE fractions were supplemented with 0.3 mg/ml insulin, dialyzed and applied to a Mono S PC 1.6/5 column and developed with a linear gradient from 0.2 M to 0.7 M KCl. jmlim@berkeley.edu 21-Dec-2011: Contacted by JYOTI. Only protein-protein interactions mouse-embryonic_stem cell Mus musculus embryonic stem cell es cells embryonic stem cells ch-ip chromatin immunoprecipitation assay ch-ip primer specific pcr primer specific pcr Fong Y.W., Inouye C., Yamaguchi T., Cattoglio C., Grubisic I., Tjian R. Nuclear extracts were prepared from NT2 cells, precipitated with ammonium sulfate and resuspended. Soluble fraction was applied to a P11-phosphocellulose column, washed and eluted at 1.0 M KCl (P1M). This P1M fraction was adjusted to 0.05% NP-40 and 10 mM imidazole, applied to a Ni-NTA resin The final Ni-FT fraction was dialyzed and applied to a Poros 20 HQ column and developed with a linear gradient from 0.52 M to 1.0 M KCl. Transcriptionally active fractions were pooled, dialyzed t applied to a Poros 20 HS column, and subjected to a linear gradient from 0.42 M to 1.0 M KCl. Active HS fractions were pooled and applied directly to hydroxyapatite (HAP) type II ceramic resin, and subjected to a linear gradient from 0.19 M to 0.5 M KPi. Active HAP fractions were pooled and separated on a Superose 6 XK 16/70 gel filtration column . Active Superose 6 fractions with an apparent molecular mass of 500-700kDa were pooled and supplemented with 0.25 mg/ml insulin. Pooled fractions were applied to a Poros 20 HE column, and developed with a linear gradient from 0.36 M to 0.86 M KCl. Active HE fractions were supplemented with 0.3 mg/ml insulin, dialyzed and applied to a Mono S PC 1.6/5 column and developed with a linear gradient from 0.2 M to 0.7 M KCl. jmlim@berkeley.edu Accepted 2011-NOV-03 by MGT 21-Dec-2011: Contacted by JYOTI. Cell (0092-8674) 2011 Only protein-protein interactions imex curation A DNA repair complex functions as an oct4/sox2 coactivator in embryonic stem cells. A DNA repair complex functions as an oct4/sox2 coactivator in embryonic stem cells. Cell (0092-8674) 2011 imex curation Fong Y.W., Inouye C., Yamaguchi T., Cattoglio C., Grubisic I., Tjian R. Nuclear extracts were prepared from NT2 cells, precipitated with ammonium sulfate and resuspended. Soluble fraction was applied to a P11-phosphocellulose column, washed and eluted at 1.0 M KCl (P1M). This P1M fraction was adjusted to 0.05% NP-40 and 10 mM imidazole, applied to a Ni-NTA resin The final Ni-FT fraction was dialyzed and applied to a Poros 20 HQ column and developed with a linear gradient from 0.52 M to 1.0 M KCl. Transcriptionally active fractions were pooled, dialyzed t applied to a Poros 20 HS column, and subjected to a linear gradient from 0.42 M to 1.0 M KCl. Active HS fractions were pooled and applied directly to hydroxyapatite (HAP) type II ceramic resin, and subjected to a linear gradient from 0.19 M to 0.5 M KPi. Active HAP fractions were pooled and separated on a Superose 6 XK 16/70 gel filtration column . Active Superose 6 fractions with an apparent molecular mass of 500-700kDa were pooled and supplemented with 0.25 mg/ml insulin. Pooled fractions were applied to a Poros 20 HE column, and developed with a linear gradient from 0.36 M to 0.86 M KCl. Active HE fractions were supplemented with 0.3 mg/ml insulin, dialyzed and applied to a Mono S PC 1.6/5 column and developed with a linear gradient from 0.2 M to 0.7 M KCl. jmlim@berkeley.edu 21-Dec-2011: Contacted by JYOTI. Only protein-protein interactions human-nt2 Homo sapiens neuronal precursor cells nt2 NT2 cells-human neuronal precursor cell line Source of these cells was Stratagene. They were derived from a lung metastasis of a 22 year old Caucasian male with a testicular germ cell tumor. Transplantation into nude mice resulted in a tumor that contained embryonal carcinoma cells. The Ntera-2 (NT2) line was established from this tumor.(taken from the Stratagene info on the web). chromatography chromatography technology column chromatography chromatography sequence tag sequence tag identification sequence tag Fong Y.W., Inouye C., Yamaguchi T., Cattoglio C., Grubisic I., Tjian R. Nuclear extracts were prepared from NT2 cells, precipitated with ammonium sulfate and resuspended. Soluble fraction was applied to a P11-phosphocellulose column, washed and eluted at 1.0 M KCl (P1M). This P1M fraction was adjusted to 0.05% NP-40 and 10 mM imidazole, applied to a Ni-NTA resin The final Ni-FT fraction was dialyzed and applied to a Poros 20 HQ column and developed with a linear gradient from 0.52 M to 1.0 M KCl. Transcriptionally active fractions were pooled, dialyzed t applied to a Poros 20 HS column, and subjected to a linear gradient from 0.42 M to 1.0 M KCl. Active HS fractions were pooled and applied directly to hydroxyapatite (HAP) type II ceramic resin, and subjected to a linear gradient from 0.19 M to 0.5 M KPi. Active HAP fractions were pooled and separated on a Superose 6 XK 16/70 gel filtration column . Active Superose 6 fractions with an apparent molecular mass of 500-700kDa were pooled and supplemented with 0.25 mg/ml insulin. Pooled fractions were applied to a Poros 20 HE column, and developed with a linear gradient from 0.36 M to 0.86 M KCl. Active HE fractions were supplemented with 0.3 mg/ml insulin, dialyzed and applied to a Mono S PC 1.6/5 column and developed with a linear gradient from 0.2 M to 0.7 M KCl. jmlim@berkeley.edu Accepted 2011-NOV-03 by MGT Cell (0092-8674) 2011 Only protein-protein interactions imex curation A DNA repair complex functions as an oct4/sox2 coactivator in embryonic stem cells. A DNA repair complex functions as an oct4/sox2 coactivator in embryonic stem cells. Cell (0092-8674) 2011 imex curation Fong Y.W., Inouye C., Yamaguchi T., Cattoglio C., Grubisic I., Tjian R. Nuclear extracts were prepared from NT2 cells, precipitated with ammonium sulfate and resuspended. Soluble fraction was applied to a P11-phosphocellulose column, washed and eluted at 1.0 M KCl (P1M). This P1M fraction was adjusted to 0.05% NP-40 and 10 mM imidazole, applied to a Ni-NTA resin The final Ni-FT fraction was dialyzed and applied to a Poros 20 HQ column and developed with a linear gradient from 0.52 M to 1.0 M KCl. Transcriptionally active fractions were pooled, dialyzed t applied to a Poros 20 HS column, and subjected to a linear gradient from 0.42 M to 1.0 M KCl. Active HS fractions were pooled and applied directly to hydroxyapatite (HAP) type II ceramic resin, and subjected to a linear gradient from 0.19 M to 0.5 M KPi. Active HAP fractions were pooled and separated on a Superose 6 XK 16/70 gel filtration column . Active Superose 6 fractions with an apparent molecular mass of 500-700kDa were pooled and supplemented with 0.25 mg/ml insulin. Pooled fractions were applied to a Poros 20 HE column, and developed with a linear gradient from 0.36 M to 0.86 M KCl. Active HE fractions were supplemented with 0.3 mg/ml insulin, dialyzed and applied to a Mono S PC 1.6/5 column and developed with a linear gradient from 0.2 M to 0.7 M KCl. jmlim@berkeley.edu 21-Dec-2011: Contacted by JYOTI. Only protein-protein interactions spofr-sf_9 Spodoptera frugiperda insect cells Fall armyworm sf_9 insect cells Derived from pupal ovarian tissue of the fall armyworm Cloned from the parent cell line, IPLB-SF-21-AE (SF-21) derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae cells are susceptible to Baculovirus infection affinity chrom affinity chromatography technology Affinity purification Affinity purification affinity chrom weight silver stain molecular weight estimation by silver staining weight silver stain Fong Y.W., Inouye C., Yamaguchi T., Cattoglio C., Grubisic I., Tjian R. Only silver staining but I think its OK as they run the purified proteins, which have been ID'd by mass spec and western on the same gel, and its a very clean purification. Nuclear extracts were prepared from NT2 cells, precipitated with ammonium sulfate and resuspended. Soluble fraction was applied to a P11-phosphocellulose column, washed and eluted at 1.0 M KCl (P1M). This P1M fraction was adjusted to 0.05% NP-40 and 10 mM imidazole, applied to a Ni-NTA resin The final Ni-FT fraction was dialyzed and applied to a Poros 20 HQ column and developed with a linear gradient from 0.52 M to 1.0 M KCl. Transcriptionally active fractions were pooled, dialyzed t applied to a Poros 20 HS column, and subjected to a linear gradient from 0.42 M to 1.0 M KCl. Active HS fractions were pooled and applied directly to hydroxyapatite (HAP) type II ceramic resin, and subjected to a linear gradient from 0.19 M to 0.5 M KPi. Active HAP fractions were pooled and separated on a Superose 6 XK 16/70 gel filtration column . Active Superose 6 fractions with an apparent molecular mass of 500-700kDa were pooled and supplemented with 0.25 mg/ml insulin. Pooled fractions were applied to a Poros 20 HE column, and developed with a linear gradient from 0.36 M to 0.86 M KCl. Active HE fractions were supplemented with 0.3 mg/ml insulin, dialyzed and applied to a Mono S PC 1.6/5 column and developed with a linear gradient from 0.2 M to 0.7 M KCl. jmlim@berkeley.edu Accepted 2011-NOV-03 by MGT 21-Dec-2011: Contacted by JYOTI. Cell (0092-8674) 2011 Only protein-protein interactions imex curation cetn2_human Centrin-2 CETN2 CALT CEN2 Caltractin isoform 1 protein protein human Homo sapiens Human MASNFKKANMASSSQRKRMSPKPELTEEQKQEIREAFDLFDADGTGTIDVKELKVAMRALGFEPKKEEIKKMISEIDKEGTGKMNFGDFLTVMTQKMSEKDTKEEILKAFKLFDDDETGKISFKNLKRVAKELGENLTDEELQEMIDEADRDGDGEVSEQEFLRIMKKTSLY 59CFD706AD7011B5 po5f1_mouse_gene POU domain, class 5, transcription factor 1 gene gene mouse Mus musculus Mouse xpc_mouse DNA repair protein complementing XP-C cells homolog Xpc Xeroderma pigmentosum group C-complementing protein homolog p125 protein protein mouse Mus musculus Mouse MAPKRTADGRRRKRGQKTEDNKVARHEESVADDFEDEKQKPRRKSSFPKVSQGKRKRGCSDPGDPTNGAAKKKVAKATAKSKNLKVLKEEALSDGDDFRDSPADCKKAKKHPKSKVVDQGTDEDDSEDDWEEVEELTEPVLDMGENSATSPSDMPVKAVEIEIETPQQAKERERSEKIKMEFETYLRRMMKRFNKEVQENMHKVHLLCLLASGFYRNSICRQPDLLAIGLSIIPIRFTKVPLQDRDAYYLSNLVKWFIGTFTVNADLSASEQDDLQTTLERRIAIYSARDNEELVHIFLLILRALQLLTRLVLSLQPIPLKSAVTKGRKSSKETSVEGPGGSSELSSNSPESHNKPTTSRRIKEEETLSEGRGKATARGKRGTGTAGSRQRRKPSCSEGEEAEQKVQGRPHARKRRVAAKVSYKEESESDGAGSGSDFEPSSGEGQHSSDEDCEPGPRKQKRASAPQRTKAGSKSASKTQRGSQCEPSSFPEASSSSSGCKRGKKVSSGAEEMADRKPAGVDQWLEVYCEPQAKWVCVDCVHGVVGQPVACYKYATKPMTYVVGIDSDGWVRDVTQRYDPAWMTATRKCRVDAEWWAETLRPYRSLLTEREKKEDQEFQAKHLDQPLPTSISTYKNHPLYALKRHLLKFQAIYPETAAVLGYCRGEAVYSRDCVHTLHSRDTWLKQARVVRLGEVPYKMVKGFSNRARKARLSEPQLHDHNDLGLYGHWQTEEYQPPIAVDGKVPRNEFGNVYLFLPSMMPVGCVQMTLPNLNRVARKLGIDCVQAITGFDFHGGYCHPVTDGYIVCEEFRDVLLAAWENEQAIIEKKEKEKKEKRALGNWKLLVRGLLIRERLKLRYGAKSEAAAPHAAGGGLSSDEEEGTSSQAEAARVLAASWPQNREDPEQKSEYTKMTRKRRAAEASHLFPFEKL 0C469AB21B4E4EE9 cetn2_mouse Centrin-2 Cetn2 Calt Caltractin isoform 1 protein protein mouse Mus musculus Mouse MASNFKKTTMASSAQRKRMSPKPELTEDQKQEIREAFDLFDADGTGTIDIKELKVAMRALGFEPKKEEIKKMISEIDKEGTGKMNFSDFLTVMTQKMSEKDTKEEILKAFKLFDDDETGKISFKNLKRVAKELGENLTDEELQEMIDEADRDGDGEVNEQEFLRIMKKTSLY 75311C8386861567 sox2_mouse Transcription factor SOX-2 Sox2 Sox-2 protein protein mouse Mus musculus Mouse MYNMMETELKPPGPQQASGGGGGGGNATAAATGGNQKNSPDRVKRPMNAFMVWSRGQRRKMAQENPKMHNSEISKRLGAEWKLLSETEKRPFIDEAKRLRALHMKEHPDYKYRPRRKTKTLMKKDKYTLPGGLLAPGGNSMASGVGVGAGLGGGLNQRMDSYAHMNGWSNGSYSMMQEQLGYPQHPGLNAHGAAQMQPMHRYVVSALQYNSMTSSQTYMNGSPTYSMSYSQQGTPGMALGSMGSVVKSEASSSPPVVTSSSHSRAPCQAGDLRDMISMYLPGAEVPEPAAPSRLHMAQHYQSGPVPGTAKYGTLPLSHM DIP protein P48432 original sequence version: 95 DIP protein P48432 original sequence version: 108 C40DE49E524C49F1 xpc_human DNA repair protein complementing XP-C cells Xeroderma pigmentosum group C-complementing protein p125 XPC XPCC protein protein human Homo sapiens Human MARKRAAGGEPRGRELRSQKSKAKSKARREEEEEDAFEDEKPPKKSLLSKVSQGKRKRGCSHPGGSADGPAKKKVAKVTVKSENLKVIKDEALSDGDDLRDFPSDLKKAHHLKRGATMNEDSNEEEEESENDWEEVEELSEPVLGDVRESTAFSRSLLPVKPVEIEIETPEQAKTRERSEKIKLEFETYLRRAMKRFNKGVHEDTHKVHLLCLLANGFYRNNICSQPDLHAIGLSIIPARFTRVLPRDVDTYYLSNLVKWFIGTFTVNAELSASEQDNLQTTLERRFAIYSARDDEELVHIFLLILRALQLLTRLVLSLQPIPLKSATAKGKKPSKERLTADPGGSSETSSQVLENHTKPKTSKGTKQEETFAKGTCRPSAKGKRNKGGRKKRSKPSSSEEDEGPGDKQEKATQRRPHGRERRVASRVSYKEESGSDEAGSGSDFELSSGEASDPSDEDSEPGPPKQRKAPAPQRTKAGSKSASRTHRGSHRKDPSLPAASSSSSSSKRGKKMCSDGEKAEKRSIAGIDQWLEVFCEQEEKWVCVDCVHGVVGQPLTCYKYATKPMTYVVGIDSDGWVRDVTQRYDPVWMTVTRKCRVDAEWWAETLRPYQSPFMDREKKEDLEFQAKHMDQPLPTAIGLYKNHPLYALKRHLLKYEAIYPETAAILGYCRGEAVYSRDCVHTLHSRDTWLKKARVVRLGEVPYKMVKGFSNRARKARLAEPQLREENDLGLFGYWQTEEYQPPVAVDGKVPRNEFGNVYLFLPSMMPIGCVQLNLPNLHRVARKLDIDCVQAITGFDFHGGYSHPVTDGYIVCEEFKDVLLTAWENEQAVIERKEKEKKEKRALGNWKLLAKGLLIRERLKRRYGPKSEAAAPHTDAGGGLSSDEEEGTSSQAEAARILAASWPQNREDEEKQKLKGGPKKTKREKKAAASHLFPFEQL 2F8C80D43FAA1256 nanog_mouse_gene gene gene mouse Mus musculus Mouse rd23b_human UV excision repair protein RAD23 homolog B XP-C repair-complementing complex 58 kDa protein RAD23B protein protein human Homo sapiens Human MQVTLKTLQQQTFKIDIDPEETVKALKEKIESEKGKDAFPVAGQKLIYAGKILNDDTALKEYKIDEKNFVVVMVTKPKAVSTPAPATTQQSAPASTTAVTSSTTTTVAQAPTPVPALAPTSTPASITPASATASSEPAPASAAKQEKPAEKPAETPVATSPTATDSTSGDSSRSNLFEDATSALVTGQSYENMVTEIMSMGYEREQVIAALRASFNNPDRAVEYLLMGIPGDRESQAVVDPPQAASTGAPQSSAVAAAAATTTATTTTTSSGGHPLEFLRNQPQFQQMRQIIQQNPSLLPALLQQIGRENPQLLQQISQHQEHFIQMLNEPVQEAGGQGGGGGGGSGGIAEAGSGHMNYIQVTPQEKEAIERLKALGFPEGLVIQAYFACEKNENLAANFLLQQNFDED C026C78273BCB289 rd23b_mouse UV excision repair protein RAD23 homolog B XP-C repair-complementing complex 58 kDa protein Rad23b Mhr23b protein protein mouse Mus musculus Mouse MQVTLKTLQQQTFKIDIDPEETVKALKEKIESEKGKDAFPVAGQKLIYAGKILSDDTALKEYKIDEKNFVVVMVTKPKAVTTAVPATTQPSSTPSPTAVSSSPAVAAAQAPAPTPALPPTSTPASTAPASTTASSEPAPAGATQPEKPAEKPAQTPVLTSPAPADSTPGDSSRSNLFEDATSALVTGQSYENMVTEIMSMGYEREQVIAALRASFNNPDRAVEYLLMGIPGDRESQAVVDPPPQAVSTGTPQSPAVAAAAATTTATTTTTSGGHPLEFLRNQPQFQQMRQIIQQNPSLLPALLQQIGRENPQLLQQISQHQEHFIQMLNEPVQEAGSQGGGGGGGGGGGGGGGGGIAEAGSGHMNYIQVTPQEKEAIERLKALGFPEGLVIQAYFACEKNENLAANFLLQQNFDED 7440E6A9C8ADF454 po5f1_mouse POU domain, class 5, transcription factor 1 Pou5f1 Oct-3 Oct-4 Otf-3 Otf3 Octamer-binding transcription factor 3 NF-A3 Octamer-binding protein 3 Octamer-binding protein 4 protein protein mouse Mus musculus Mouse MAGHLASDFAFSPPPGGGDGSAGLEPGWVDPRTWLSFQGPPGGPGIGPGSEVLGISPCPPAYEFCGGMAYCGPQVGLGLVPQVGVETLQPEGQAGARVESNSEGTSSEPCADRPNAVKLEKVEPTPEESQDMKALQKELEQFAKLLKQKRITLGYTQADVGLTLGVLFGKVFSQTTICRFEALQLSLKNMCKLRPLLEKWVEEADNNENLQEICKSETLVQARKRKRTSIENRVRWSLETMFLKCPKPSLQQITHIANQLGLEKDVVRVWFCNRRQKGKRSSIEYSQREEYEATGTPFPGGAVSFPLPPGPHFGTPGYGSPHFTTLYSVPFPEGEAFPSVPVTALGSPMHSN DIP protein P20263 original sequence version: 87 DIP protein P20263 original sequence version: 121 DIP protein P20263 original sequence version: 118 DIP protein P20263 original sequence version: 137 DIP protein P20263 original sequence version: 107 757E41DF52286714 cetn2-rad23b 6 8 unspecified role unspecified role neutral component neutral component 15 unspecified role unspecified role neutral component neutral component 13 unspecified role unspecified role neutral component neutral component association association Fig. 3A cetn2-rad23b-1 3 13 unspecified role unspecified role neutral component neutral component 15 unspecified role unspecified role neutral component neutral component 8 unspecified role unspecified role neutral component neutral component association association Fig. 3B rad23b-cetn2 7 13 unspecified role unspecified role prey prey region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 15 unspecified role unspecified role bait bait region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 8 unspecified role unspecified role prey prey association association Fig. 4A rad23b-cetn2-2 1 17 unspecified role unspecified role prey prey 10 unspecified role unspecified role prey prey 16 unspecified role unspecified role bait bait 12 unspecified role unspecified role prey prey 11 unspecified role unspecified role prey prey association association Fig. 4D rad23b-cetn2-3 2 8 unspecified role unspecified role prey prey 13 unspecified role unspecified role prey prey region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 15 unspecified role unspecified role bait bait region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined association association Fig. 4E rad23b-cetn2-4 2 15 unspecified role unspecified role bait bait region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 13 unspecified role unspecified role prey prey region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain physical association physical association Fig. 4E, Supp Fig. 3C xpc-cetn2 4 8 unspecified role unspecified role prey prey 13 unspecified role unspecified role bait bait region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association Fig. 4E rad23b-nanog 5 16 unspecified role unspecified role bait bait 14 unspecified role unspecified role prey prey distal enhancer binding region binding-associated region binding region undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined transcription start site binding region binding-associated region binding region undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined association association Fig. 6A Analysis revealed that RAD23B occupancy sites coincide with those reported for Oct4 and Sox2 rad23b-po5f1 5 9 unspecified role unspecified role prey prey transcription start site binding region binding-associated region binding region undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined distal enhancer binding region binding-associated region binding region undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 16 unspecified role unspecified role bait bait association association Fig. 6A Analysis revealed that RAD23B occupancy sites coincide with those reported for Oct4 and Sox2 sox2-nanog 5 14 unspecified role unspecified role prey prey distal enhancer binding region binding-associated region binding region undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined distal enhancer binding region binding-associated region binding region undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 12 unspecified role unspecified role bait bait association association Supp. Fig. 5A Analysis revealed that RAD23B occupancy sites coincide with those reported for Oct4 and Sox2 sox2-po5f1 5 9 unspecified role unspecified role prey prey distal enhancer binding region binding-associated region binding region undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 12 unspecified role unspecified role bait bait association association Fig. S5A Analysis revealed that RAD23B occupancy sites coincide with those reported for Oct4 and Sox2 rad23b-cetn2-5 1 16 unspecified role unspecified role prey prey 10 unspecified role unspecified role prey prey 11 unspecified role unspecified role prey prey 17 unspecified role unspecified role bait bait association association Fig. 4C