Figure 5. SET7/9-mediated YY1 methylation is involved in YY1-regulated gene transcription.

(A) HeLa cells were transfected with control siRNA or siRNA specifically targeting YY1, followed by immunoblotting using antibodies as indicated to examine the knock-down efficiency of siYY1. (B) Gro-seq experiments were performed with nuclei collected from cells described in (A). Genes regulated by YY1 were displayed using pie chart (top, P < 0.001). Among all genes regulated by YY1, those with or without YY1 binding on promoter regions were displayed using Venn diagram (bottom two). (C) Gene ontology analysis was performed for genes positively-regulated by YY1 as shown in (B) using DAVID. Top ten enriched gene ontology (GO) terms were shown. (D) Knock-down efficiency of siSET7/9 was examined through immunoblotting using antibodies as indicated. (E) HeLa cells were transfected with control siRNA or siRNA specifically targeting YY1 or SET7/9, followed by RT-qPCR analysis to examine mRNA levels of selected genes as indicated. Data shown was the relative fold change compared to control samples after normalization to actin. Experiments were repeated three times and representative data was shown. (F) HeLa cells were transfected with control vector or vectors expressing YY1(wt), YY1(K173R) or YY1(K411R), followed by RT-qPCR analysis to examine mRNA levels of selected genes as indicated. Data shown was the relative fold change compared to control samples after normalization to actin (±s.e.m., *P < 0.05, **P < 0.01, ***P < 0.001). Experiments were repeated four times and representative data was shown.