Fig. 7

RA treatment notably enhanced binding of not only RARs but also RIPK1 and RIPK3 to RAREs of RA-inducible genes in the absence of Casp8 expression. a Tet-On shCasp8 ES cells were cultured with or without 1 μg/ml Dox for 4 days and then treated with or without 1 μM RA for 24 h in the presence or absence of Dox. Subsequently, ChIP analysis for the Rarb-specific RARE or Gapdh promoter region using an anti-RIPK1 antibody was carried out. Tet-On shCasp8 P19 cells expressing shRipk3 (shRipk3 + ) or shLacZ (shRipk3-) together with or without the expression of shRipk3-resistant 3xFlag-Wt RIPK3, kinase-negative 551 A mutant RIPK3, or RHIM AAAA mutant RIPK3 were treated with or without 1 μM RA for 24 h in the presence or absence of 1 μg/ml Dox, and subjected to ChIP analysis for the Cyr26a1-specific RARE or Gapdh promoter region using anti-Flag antibody or control IgG (b), or using anti-RAR antibody or control IgG (c). Vector, an empty vector. d Dox (1 μg/ml)-treated Tet-On shCasp8 ES cells expressing shRipk3 and shRipk3-resistant 3xFlag-Wt RIPK3 were subjected to immunoprecipitation (IP) after 4 days formation of EBs. EBs were treated with 1 μM RA for last 2 days. Immunoprecipitates with control IgG and ant-Flag antibodies were subjected to western blot analysis using indicated antibodies. Molecular weight markers are indicated (kDa). **p < 0.01 and *p < 0.05