Fig 1: C2EIP regulates PGC generation by activating the HH signaling pathway.a Expression of PTCH2 protein in ESC, PGC, and SSC was tested by western Blot. b The phosphorylation level of PTCH2 was revealed by co-IP in ESC, PGC, and SSC before and after C2EIP knockout, as well as in the 0-day and 4-day RA induction in vitro model. The phosphorylation level of PTCH2 in ESC was the control. c, d, e The PTCH2 ubiquitylation level was revealed by co-IPin ESC, PGC, and SSC before and after C2EIP knockout as well as in the 0-day and 4-day RA in vitro induction test
Fig 2: C2EIP and PTCH2 are co-expressed at the intracellular membrane.a Interaction between PTCH2 and C2EIP proteins was observed by GSTpull-down test. IgG antibody was used as a negative control. b Indirect immunofluorescence results indicate that PTCH2 protein is localized in cell membranes, Scale bar: 30 µm. SSEA-1was used as acell membrane marker. c Indirect immunofluorescence results indicate that PTCH2 and C2EIP colocalize at cell membranes, Scale bar: 20 µm. d Co-immunoprecipitation from PGC detected an interaction between GST-C2EIP and PTCH2 proteins. IgG antibody was used as negative control. e Western blot analysis showed that GST-C2EIP and PTCH2 are expressed in the cytoplasm of PGC
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