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 Suppression of estrogen receptor transcriptional activity by connective tissue growth factor. Suppression of estrogen receptor transcriptional activity by connective tissue growth factor. PloS one 2010 imex curation Cheng L., Yang Z., Wang X., Jiao Y., Xie X., Lin J., Zhang H., Han J., Jiang K., Ye Q., Cheng L., Yang Z., Wang X., Jiao Y., Xie X., Lin J., Zhang H., Han J., Jiang K., Ye Q., Cheng L., Yang Z., Wang X., Jiao Y., Xie X., Lin J., Zhang H., Han J., Jiang K., Ye Q., Cheng L., Yang Z., Wang X., Jiao Y., Xie X., Lin J., Zhang H., Han J., Jiang K., Ye Q. I have checked the references, ERalpha and beta are human. I did not manage to get the IMEx ID from IMEx Central (it doesn't work). yeqn66@yahoo.com 09-May-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation human-mcf7 Homo sapiens mammary carcinoma cells mcf7 mammary carcinoma cells fluorescence imaging fluorescence microscopy fluorescence imaging immunostaining immunostaining Cheng L., Yang Z., Wang X., Jiao Y., Xie X., Lin J., Zhang H., Han J., Jiang K., Ye Q., Cheng L., Yang Z., Wang X., Jiao Y., Xie X., Lin J., Zhang H., Han J., Jiang K., Ye Q., Cheng L., Yang Z., Wang X., Jiao Y., Xie X., Lin J., Zhang H., Han J., Jiang K., Ye Q., Cheng L., Yang Z., Wang X., Jiao Y., Xie X., Lin J., Zhang H., Han J., Jiang K., Ye Q. I have checked the references, ERalpha and beta are human. I did not manage to get the IMEx ID from IMEx Central (it doesn't work). yeqn66@yahoo.com Accepted 2018-OCT-16 AT 10:59 BST AT 10:59 BST by LIVIAP PloS one 2010 imex curation Aggiunto! Suppression of estrogen receptor transcriptional activity by connective tissue growth factor. Suppression of estrogen receptor transcriptional activity by connective tissue growth factor. PloS one 2010 imex curation Cheng L., Yang Z., Wang X., Jiao Y., Xie X., Lin J., Zhang H., Han J., Jiang K., Ye Q., Cheng L., Yang Z., Wang X., Jiao Y., Xie X., Lin J., Zhang H., Han J., Jiang K., Ye Q., Cheng L., Yang Z., Wang X., Jiao Y., Xie X., Lin J., Zhang H., Han J., Jiang K., Ye Q., Cheng L., Yang Z., Wang X., Jiao Y., Xie X., Lin J., Zhang H., Han J., Jiang K., Ye Q. I have checked the references, ERalpha and beta are human. I did not manage to get the IMEx ID from IMEx Central (it doesn't work). yeqn66@yahoo.com 09-May-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro pull down pull down anti tag western anti tag western blot anti tag western Cheng L., Yang Z., Wang X., Jiao Y., Xie X., Lin J., Zhang H., Han J., Jiang K., Ye Q., Cheng L., Yang Z., Wang X., Jiao Y., Xie X., Lin J., Zhang H., Han J., Jiang K., Ye Q., Cheng L., Yang Z., Wang X., Jiao Y., Xie X., Lin J., Zhang H., Han J., Jiang K., Ye Q., Cheng L., Yang Z., Wang X., Jiao Y., Xie X., Lin J., Zhang H., Han J., Jiang K., Ye Q. I have checked the references, ERalpha and beta are human. I did not manage to get the IMEx ID from IMEx Central (it doesn't work). yeqn66@yahoo.com Accepted 2018-OCT-16 AT 10:59 BST AT 10:59 BST by LIVIAP PloS one 2010 imex curation Aggiunto! Suppression of estrogen receptor transcriptional activity by connective tissue growth factor. Suppression of estrogen receptor transcriptional activity by connective tissue growth factor. PloS one 2010 imex curation Cheng L., Yang Z., Wang X., Jiao Y., Xie X., Lin J., Zhang H., Han J., Jiang K., Ye Q., Cheng L., Yang Z., Wang X., Jiao Y., Xie X., Lin J., Zhang H., Han J., Jiang K., Ye Q., Cheng L., Yang Z., Wang X., Jiao Y., Xie X., Lin J., Zhang H., Han J., Jiang K., Ye Q., Cheng L., Yang Z., Wang X., Jiao Y., Xie X., Lin J., Zhang H., Han J., Jiang K., Ye Q. I have checked the references, ERalpha and beta are human. I did not manage to get the IMEx ID from IMEx Central (it doesn't work). yeqn66@yahoo.com 09-May-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast 2 hybrid two hybrid 2H 2h Gal4 transcription regeneration two-hybrid yeast two hybrid 2-hybrid classical two hybrid classical two hybrid 2H two-hybrid 2-hybrid Gal4 transcription regeneration 2h yeast two hybrid Y2H Y-2H 2 hybrid nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence Cheng L., Yang Z., Wang X., Jiao Y., Xie X., Lin J., Zhang H., Han J., Jiang K., Ye Q., Cheng L., Yang Z., Wang X., Jiao Y., Xie X., Lin J., Zhang H., Han J., Jiang K., Ye Q., Cheng L., Yang Z., Wang X., Jiao Y., Xie X., Lin J., Zhang H., Han J., Jiang K., Ye Q., Cheng L., Yang Z., Wang X., Jiao Y., Xie X., Lin J., Zhang H., Han J., Jiang K., Ye Q. I have checked the references, ERalpha and beta are human. I did not manage to get the IMEx ID from IMEx Central (it doesn't work). yeqn66@yahoo.com Accepted 2018-OCT-16 AT 10:59 BST AT 10:59 BST by LIVIAP human mammary cDNA library PloS one 2010 imex curation Aggiunto! Suppression of estrogen receptor transcriptional activity by connective tissue growth factor. Suppression of estrogen receptor transcriptional activity by connective tissue growth factor. PloS one 2010 imex curation Cheng L., Yang Z., Wang X., Jiao Y., Xie X., Lin J., Zhang H., Han J., Jiang K., Ye Q., Cheng L., Yang Z., Wang X., Jiao Y., Xie X., Lin J., Zhang H., Han J., Jiang K., Ye Q., Cheng L., Yang Z., Wang X., Jiao Y., Xie X., Lin J., Zhang H., Han J., Jiang K., Ye Q., Cheng L., Yang Z., Wang X., Jiao Y., Xie X., Lin J., Zhang H., Han J., Jiang K., Ye Q. I have checked the references, ERalpha and beta are human. I did not manage to get the IMEx ID from IMEx Central (it doesn't work). yeqn66@yahoo.com 09-May-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast 2 hybrid two hybrid 2H 2h Gal4 transcription regeneration two-hybrid yeast two hybrid 2-hybrid classical two hybrid classical two hybrid 2H two-hybrid 2-hybrid Gal4 transcription regeneration 2h yeast two hybrid Y2H Y-2H 2 hybrid predetermined predetermined participant predetermined Cheng L., Yang Z., Wang X., Jiao Y., Xie X., Lin J., Zhang H., Han J., Jiang K., Ye Q., Cheng L., Yang Z., Wang X., Jiao Y., Xie X., Lin J., Zhang H., Han J., Jiang K., Ye Q., Cheng L., Yang Z., Wang X., Jiao Y., Xie X., Lin J., Zhang H., Han J., Jiang K., Ye Q., Cheng L., Yang Z., Wang X., Jiao Y., Xie X., Lin J., Zhang H., Han J., Jiang K., Ye Q. I have checked the references, ERalpha and beta are human. I did not manage to get the IMEx ID from IMEx Central (it doesn't work). yeqn66@yahoo.com Accepted 2018-OCT-16 AT 10:59 BST AT 10:59 BST by LIVIAP PloS one 2010 imex curation Aggiunto! Suppression of estrogen receptor transcriptional activity by connective tissue growth factor. Suppression of estrogen receptor transcriptional activity by connective tissue growth factor. PloS one 2010 imex curation Cheng L., Yang Z., Wang X., Jiao Y., Xie X., Lin J., Zhang H., Han J., Jiang K., Ye Q., Cheng L., Yang Z., Wang X., Jiao Y., Xie X., Lin J., Zhang H., Han J., Jiang K., Ye Q., Cheng L., Yang Z., Wang X., Jiao Y., Xie X., Lin J., Zhang H., Han J., Jiang K., Ye Q., Cheng L., Yang Z., Wang X., Jiao Y., Xie X., Lin J., Zhang H., Han J., Jiang K., Ye Q. I have checked the references, ERalpha and beta are human. I did not manage to get the IMEx ID from IMEx Central (it doesn't work). yeqn66@yahoo.com 09-May-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro elisa enzyme linked immunosorbent assay ELISA ELISA elisa antibody detection identification by antibody antibody detection Cheng L., Yang Z., Wang X., Jiao Y., Xie X., Lin J., Zhang H., Han J., Jiang K., Ye Q., Cheng L., Yang Z., Wang X., Jiao Y., Xie X., Lin J., Zhang H., Han J., Jiang K., Ye Q., Cheng L., Yang Z., Wang X., Jiao Y., Xie X., Lin J., Zhang H., Han J., Jiang K., Ye Q., Cheng L., Yang Z., Wang X., Jiao Y., Xie X., Lin J., Zhang H., Han J., Jiang K., Ye Q. I have checked the references, ERalpha and beta are human. I did not manage to get the IMEx ID from IMEx Central (it doesn't work). yeqn66@yahoo.com Accepted 2018-OCT-16 AT 10:59 BST AT 10:59 BST by LIVIAP PloS one 2010 imex curation Aggiunto! Suppression of estrogen receptor transcriptional activity by connective tissue growth factor. Suppression of estrogen receptor transcriptional activity by connective tissue growth factor. PloS one 2010 imex curation Cheng L., Yang Z., Wang X., Jiao Y., Xie X., Lin J., Zhang H., Han J., Jiang K., Ye Q., Cheng L., Yang Z., Wang X., Jiao Y., Xie X., Lin J., Zhang H., Han J., Jiang K., Ye Q., Cheng L., Yang Z., Wang X., Jiao Y., Xie X., Lin J., Zhang H., Han J., Jiang K., Ye Q., Cheng L., Yang Z., Wang X., Jiao Y., Xie X., Lin J., Zhang H., Han J., Jiang K., Ye Q. I have checked the references, ERalpha and beta are human. I did not manage to get the IMEx ID from IMEx Central (it doesn't work). yeqn66@yahoo.com 09-May-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation human-mcf7 Homo sapiens mammary carcinoma cells mcf7 mammary carcinoma cells anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Cheng L., Yang Z., Wang X., Jiao Y., Xie X., Lin J., Zhang H., Han J., Jiang K., Ye Q., Cheng L., Yang Z., Wang X., Jiao Y., Xie X., Lin J., Zhang H., Han J., Jiang K., Ye Q., Cheng L., Yang Z., Wang X., Jiao Y., Xie X., Lin J., Zhang H., Han J., Jiang K., Ye Q., Cheng L., Yang Z., Wang X., Jiao Y., Xie X., Lin J., Zhang H., Han J., Jiang K., Ye Q. I have checked the references, ERalpha and beta are human. I did not manage to get the IMEx ID from IMEx Central (it doesn't work). yeqn66@yahoo.com Accepted 2018-OCT-16 AT 10:59 BST AT 10:59 BST by LIVIAP PloS one 2010 imex curation Aggiunto! esr2_human Estrogen receptor beta ESR2 ESTRB NR3A2 Nuclear receptor subfamily 3 group A member 2 protein protein human Homo sapiens Human MDIKNSPSSLNSPSSYNCSQSILPLEHGSIYIPSSYVDSHHEYPAMTFYSPAVMNYSIPSNVTNLEGGPGRQTTSPNVLWPTPGHLSPLVVHRQLSHLYAEPQKSPWCEARSLEHTLPVNRETLKRKVSGNRCASPVTGPGSKRDAHFCAVCSDYASGYHYGVWSCEGCKAFFKRSIQGHNDYICPATNQCTIDKNRRKSCQACRLRKCYEVGMVKCGSRRERCGYRLVRRQRSADEQLHCAGKAKRSGGHAPRVRELLLDALSPEQLVLTLLEAEPPHVLISRPSAPFTEASMMMSLTKLADKELVHMISWAKKIPGFVELSLFDQVRLLESCWMEVLMMGLMWRSIDHPGKLIFAPDLVLDRDEGKCVEGILEIFDMLLATTSRFRELKLQHKEYLCVKAMILLNSSMYPLVTATQDADSSRKLAHLLNAVTDALVWVIAKSGISSQQQSMRLANLLMLLSHVRHASNKGMEHLLNMKCKNVVPVYDLLLEMLNAHVLRGCKSSITGSECSPAEDSKSKEGSQNPQSQ DIP protein Q92731 original sequence version: 171 8CAE34215992418A ccn2_human CCN family member 2 CCN2 CTGF Cellular communication network factor 2 Connective tissue growth factor HCS24 Hypertrophic chondrocyte-specific protein 24 IGFBP8 Insulin-like growth factor-binding protein 8 protein protein human Homo sapiens Human MTAASMGPVRVAFVVLLALCSRPAVGQNCSGPCRCPDEPAPRCPAGVSLVLDGCGCCRVCAKQLGELCTERDPCDPHKGLFCHFGSPANRKIGVCTAKDGAPCIFGGTVYRSGESFQSSCKYQCTCLDGAVGCMPLCSMDVRLPSPDCPFPRRVKLPGKCCEEWVCDEPKDQTVVGPALAAYRLEDTFGPDPTMIRANCLVQTTEWSACSKTCGMGISTRVTNDNASCRLEKQSRLCMVRPCEADLEENIKKGKKCIRTPKISKPIKFELSGCTSMKTYRAKFCGVCTDGRCCTPHRTTTLPVEFKCPDGEVMKKNMMFIKTCACHYNCPGDNDIFESLYYRKMYGDMA 62CF878C7F57EC1F esr1_human Estrogen receptor ESR1 ESR NR3A1 Estradiol receptor ER-alpha Nuclear receptor subfamily 3 group A member 1 protein protein human Homo sapiens Human MTMTLHTKASGMALLHQIQGNELEPLNRPQLKIPLERPLGEVYLDSSKPAVYNYPEGAAYEFNAAAAANAQVYGQTGLPYGPGSEAAAFGSNGLGGFPPLNSVSPSPLMLLHPPPQLSPFLQPHGQQVPYYLENEPSGYTVREAGPPAFYRPNSDNRRQGGRERLASTNDKGSMAMESAKETRYCAVCNDYASGYHYGVWSCEGCKAFFKRSIQGHNDYMCPATNQCTIDKNRRKSCQACRLRKCYEVGMMKGGIRKDRRGGRMLKHKRQRDDGEGRGEVGSAGDMRAANLWPSPLMIKRSKKNSLALSLTADQMVSALLDAEPPILYSEYDPTRPFSEASMMGLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEILMIGLVWRSMEHPGKLLFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEFVCLKSIILLNSGVYTFLSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRLAQLLLILSHIRHMSNKGMEHLYSMKCKNVVPLYDLLLEMLDAHRLHAPTSRGGASVEETDQSHLATAGSTSSHSLQKYYITGEAEGFPATV DIP protein P03372 original sequence version: 214 DIP protein P03372 original sequence version: 139 DIP protein P03372 original sequence version: 167 5455C57AB0CCCAA7 ctgf-1 3 8 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell 7 predetermined predetermined participant predetermined nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell binding range binding range sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain physical association physical association nfa To identify proteins that could be involved in regulation of estrogen signaling, we screened a human mammary cDNA library using amino acids 131–324 containing the ERβ DBD and hinge domains as bait in the yeast two-hybrid system. ctgf-4 4 8 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell 7 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell binding region binding region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain physical association physical association f1a f1b TAs shown in Fig. 1A, transformation of yeast cells with CTGF and ERβ(131–324), but not with other control plasmids, activate the his (growth) and lacZ (β-gal) reporter genes. CTGF did not interact with ERβ(1–167) containing the AF1 and ERβ(286–530) containing the AF2 (Fig. 1B), suggesting the specific interaction of CTGF with ERβ(131–324). esr1-ctgf-1 4 8 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell 9 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell binding region binding region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain physical association physical association f1a f1b As shown in Fig. 1C, the ERα(180–282) containing the DBD specifically interacted with CTGF, but the ERα(1–185) containing the AF1 and the ERα(282–595) containing the hinge and AF2 regions did not. esr2-ctgf-1 6 8 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell flag tag flag tag 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 7 unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level physical association physical association f2a To further confirm the interaction between CTGF and ERα/ERβ, coimmunoprecipitation experiments were performed with MCF7 breast cancer cells. FLAG-tagged CTGF coimmunoprecipitated ERα and ERβ (Fig. 2A). ctgf-5 6 8 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell flag tag flag tag 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 9 unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level physical association physical association f2a To further confirm the interaction between CTGF and ERα/ERβ, coimmunoprecipitation experiments were performed with MCF7 breast cancer cells. FLAG-tagged CTGF coimmunoprecipitated ERα and ERβ (Fig. 2A). ctgf-6 5 8 unspecified role unspecified role bait bait purified purified 9 unspecified role unspecified role prey prey cell lysate cell lysate physiological level physiological level endogenous level endogenous endogenous endogenous level physical association physical association f2b a sensitive ELISA-based protein-protein binding detection method was employed to determine interaction of endogenous CTGF with endogenous ERα. ctgf-esr1-1 1 8 unspecified role unspecified role neutral component neutral component physiological level physiological level endogenous level endogenous endogenous endogenous level fixed cell fixed cell 9 unspecified role unspecified role neutral component neutral component physiological level physiological level endogenous level endogenous endogenous endogenous level fixed cell fixed cell colocalization colocalization f2c Moreover, immunofluorescence analysis of MCF7 cells showed that endogenous CTGF protein colocalized with endogenous ERα protein in both the cytoplasm and the membrane (Fig. 2C). ctgf-esr2-1 2 8 unspecified role unspecified role prey prey purified purified his tag his tag 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 7 unspecified role unspecified role bait bait purified purified gst tag gst tag gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined direct interaction direct interaction f2d To determine whether CTGF directly interacts with ERα and ERβ in vitro, GST pull-down experiments were performed in which purified GST-ERα or GST-ERβ was incubated with purified His-CTGF or His-Nov. As shown in Fig. 2D, CTGF, but not Nov, another CCN family member, directly interacted with ERα and ERβ. The CTGF(182–349) fragment containing the TSP-1 and CT domains bound specifically to ERα and ERβ, whereas the CTGF(27–187) fragment containing the IGFBP and VWC domains but lacking the signal peptide did not bind ERα and ERβ (Fig. 3B, left panel). esr2-ctgf-2 2 8 unspecified role unspecified role prey prey purified purified his tag his tag 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 9 unspecified role unspecified role bait bait purified purified gst tag gst tag gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined binding region binding region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain direct interaction direct interaction f2d f3a Further deletion analysis showed that the CTGF(182–250) containing the TSP-1 domain, but not the CTGF(244–349) containing the CT domain, is sufficient for ER binding (Fig. 3B, right panel). esr2-ctgf-3 2 9 unspecified role unspecified role prey prey in vitro translated in vitro translated protein in vitro translated 8 unspecified role unspecified role bait bait purified purified gst tag gst tag gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined binding region binding region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 7 unspecified role unspecified role prey prey in vitro translated in vitro translated protein in vitro translated association association f3b To determine whether CTGF directly interacts with ERα and ERβ in vitro, GST pull-down experiments were performed in which purified GST-ERα or GST-ERβ was incubated with purified His-CTGF or His-Nov. As shown in Fig. 2D, CTGF, but not Nov, another CCN family member, directly interacted with ERα and ERβ. The CTGF(182–349) fragment containing the TSP-1 and CT domains bound specifically to ERα and ERβ, whereas the CTGF(27–187) fragment containing the IGFBP and VWC domains but lacking the signal peptide did not bind ERα and ERβ (Fig. 3B, left panel). esr1-ctgf-2 2 8 unspecified role unspecified role bait bait purified purified gst tag gst tag gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined binding region binding region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 9 unspecified role unspecified role prey prey in vitro translated in vitro translated protein in vitro translated physical association physical association f3b Further deletion analysis showed that the CTGF(182–250) containing the TSP-1 domain, but not the CTGF(244–349) containing the CT domain, is sufficient for ER binding (Fig. 3B, right panel).