MINT MINT, Dpt of Biology, University of Rome Tor Vergata MINT http://mint.bio.uniroma2.it/mint url>http://mint.bio.uniroma2.it/mint/ https://mint.bio.uniroma2.it MINT-[0-9]+ https://mint.bio.uniroma2.it/index.php/results-interactions/?id=${ac} The Molecular INTeraction database (MINT) is a relational database designed to store interactions between biological molecules. http://mint.bio.uniroma2.it hCAF1/CNOT7 regulates interferon signalling by targeting STAT1. hCAF1/CNOT7 regulates interferon signalling by targeting STAT1. EMBO J. (0261-4189) 2013 imex curation Chapat C., Kolytcheff C., Le Romancer M., Auboeuf D., De La Grange P., Chettab K., Sentis S., Corbo L. laura.corbo@lyon.unicancer.fr 22-Nov-2013: Contacted by IntAct-Help. Only protein-protein interactions human-mcf7 Homo sapiens mammary carcinoma cells mcf7 mammary carcinoma cells anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Chapat C., Kolytcheff C., Le Romancer M., Auboeuf D., De La Grange P., Chettab K., Sentis S., Corbo L. laura.corbo@lyon.unicancer.fr Accepted 2013-NOV-06 by LPERFETTO 22-Nov-2013: Contacted by IntAct-Help. EMBO J. (0261-4189) 2013 Only protein-protein interactions imex curation hCAF1/CNOT7 regulates interferon signalling by targeting STAT1. hCAF1/CNOT7 regulates interferon signalling by targeting STAT1. EMBO J. (0261-4189) 2013 imex curation Chapat C., Kolytcheff C., Le Romancer M., Auboeuf D., De La Grange P., Chettab K., Sentis S., Corbo L. laura.corbo@lyon.unicancer.fr 22-Nov-2013: Contacted by IntAct-Help. Only protein-protein interactions human-mcf7 Homo sapiens mammary carcinoma cells mcf7 mammary carcinoma cells imaging technique imaging technique microscopy immunostaining immunostaining Chapat C., Kolytcheff C., Le Romancer M., Auboeuf D., De La Grange P., Chettab K., Sentis S., Corbo L. laura.corbo@lyon.unicancer.fr Accepted 2013-NOV-06 by LPERFETTO 22-Nov-2013: Contacted by IntAct-Help. EMBO J. (0261-4189) 2013 Only protein-protein interactions imex curation cnot7_human CCR4-NOT transcription complex subunit 7 CNOT7 CAF1 CCR4-associated factor 1 BTG1-binding factor 1 Caf1a protein protein human Homo sapiens Human MPAATVDHSQRICEVWACNLDEEMKKIRQVIRKYNYVAMDTEFPGVVARPIGEFRSNADYQYQLLRCNVDLLKIIQLGLTFMNEQGEYPPGTSTWQFNFKFNLTEDMYAQDSIELLTTSGIQFKKHEEEGIETQYFAELLMTSGVVLCEGVKWLSFHSGYDFGYLIKILTNSNLPEEELDFFEILRLFFPVIYDVKYLMKSCKNLKGGLQEVAEQLELERIGPQHQAGSDSLLTGMAFFKMREMFFEDHIDDAKYCGHLYGLGSGSSYVQNGTGNAYEEEANKQS DIP protein Q9UIV1 original sequence version: 95 2AF23ED27E06EFDB stat1_human Signal transducer and activator of transcription 1-alpha/beta STAT1 Transcription factor ISGF-3 components p91/p84 protein protein human Homo sapiens Human MSQWYELQQLDSKFLEQVHQLYDDSFPMEIRQYLAQWLEKQDWEHAANDVSFATIRFHDLLSQLDDQYSRFSLENNFLLQHNIRKSKRNLQDNFQEDPIQMSMIIYSCLKEERKILENAQRFNQAQSGNIQSTVMLDKQKELDSKVRNVKDKVMCIEHEIKSLEDLQDEYDFKCKTLQNREHETNGVAKSDQKQEQLLLKKMYLMLDNKRKEVVHKIIELLNVTELTQNALINDELVEWKRRQQSACIGGPPNACLDQLQNWFTIVAESLQQVRQQLKKLEELEQKYTYEHDPITKNKQVLWDRTFSLFQQLIQSSFVVERQPCMPTHPQRPLVLKTGVQFTVKLRLLVKLQELNYNLKVKVLFDKDVNERNTVKGFRKFNILGTHTKVMNMEESTNGSLAAEFRHLQLKEQKNAGTRTNEGPLIVTEELHSLSFETQLCQPGLVIDLETTSLPVVVISNVSQLPSGWASILWYNMLVAEPRNLSFFLTPPCARWAQLSEVLSWQFSSVTKRGLNVDQLNMLGEKLLGPNASPDGLIPWTRFCKENINDKNFPFWLWIESILELIKKHLLPLWNDGCIMGFISKERERALLKDQQPGTFLLRFSESSREGAITFTWVERSQNGGEPDFHAVEPYTKKELSAVTFPDIIRNYKVMAAENIPENPLKYLYPNIDKDHAFGKYYSRPKEAPEPMELDGPKGTGYIKTELISVSEVHPSRLQTTDNLLPMSPEEFDEVSRIVGSVEFDSMMNTV 054A813522364BA6 cnot7-stat1-1 1 hCAF1 3 western blot western blot Immuno blot Immuno blot experimental particp experimental participant identification experimental particp unspecified role unspecified role bait bait living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level STAT1 4 western blot western blot Immuno blot Immuno blot experimental particp experimental participant identification experimental particp unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level physical association physical association The same occurs in U937 cells. f6c sf5b cnot7-stat1-3 2 hCAF1 3 immunostaining immunostaining experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component physiological level physiological level endogenous level endogenous endogenous endogenous level fixed cell fixed cell STAT1 4 immunostaining immunostaining experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component physiological level physiological level endogenous level endogenous endogenous endogenous level fixed cell fixed cell colocalization colocalization f6d sf7 sf8 To determine in which cellular compartment this interaction occurred and whether it was regulated by IFN induction, we used an in situ proximity ligation assay (PLA), a technology capable of detecting protein interactions and the localization of interactions with high specificity and sensitivity. In the absence of IFN, the discrete spot-like signals indicated the interaction between endogenous hCAF1 and STAT1, as shown in Figure 6Da. Importantly, since the image in Figure 6Da was recorded in the fixed plane, the localization of each spot inside the cell could be unclear. By imaging cells with confocal microscopy, Z-stack projections showed unambiguously that the interaction spots were exclusively localized in the cytoplasm of resting MCF7 cells (see Supplementary Figure 7; compare a reconstitution of a three-dimensional (3D) image in Supplementary Figure 7A to a single median section cutting the nucleus, in Supplementary Figure 7B). The same occurs in U937 cells.