FIGURE 6.

ZMYND8 interacts with RNA polymerase II phospho-Ser5 in a DNA-dependent manner. A, ZMYND8 was co-immunoprecipitated from SH-SY5Y cells and immunoblotted with α-RNA polymerase II non-CTD (epitope at N terminus), α-RNA polymerase II phospho-Ser-5, and α-RNA polymerase II phospho-Ser-2 antibodies. IgG serves as negative control. B–D, similarly, RNA polymerase II non-CTD (B), RNA pol II phospho-Ser-5 (C), and RNA pol II phospho-Ser-2 (D) was co-immunoprecipitated from SH-SY5Y cells and immunoblotted with α-ZMYND8 antibody. E, co-IP of RNA pol II phospho-Ser-5 from 4 days ATRA- or DMSO-treated SH-SY5Y cells followed by immunoblotting with α-ZMYND8 antibody. IgG serves as negative control. F, RNA pol II phospho-Ser-5 was co-immunoprecipitated from SH-SY5Y cells after DNase I treatment of lysates and immunoblotted with α-ZMYND8 antibody. IgG serves as negative control. G, ChIP assays were performed after ZMYND8 siRNA or a non-targeting siRNA transfection during 2 days of ATRA or DMSO treatment in SH-SY5Y cells with α-RNA polymerase II phospho-Ser-5 antibody. Relative fold was calculated by normalizing RNA pol II phospho-Ser-5 with IgG and represented as ATRA-treated differentiated cells over DMSO-treated undifferentiated cells.