Figure 5. FAM47E regulates the expression of PRMT5 target genes.

(A, B) HEK293 cells were transfected with GFP vector or GFP-FAM47E construct or control siRNA or FAM47E siRNA or co-transfected with GFP-FAM47E construct and control siRNA or PRMT5 siRNA. After 48 h of transfections, the whole cell lysate and total RNA were isolated from these cells. (A) The cell lysates were immunoblotted and probed with GFP antibody or PRMT5 antibody or FAM47E antibody or β actin antibody. (B) The transcripts levels of the indicated PRMT5 target genes in these cells were quantified by using quantitative RT-PCR. The mRNA levels of indicated PRMT5 target genes were normalized to GAPDH expression and are presented relative to the control sample. The values represent the mean of three independent experiments, with error bars representing standard deviations. Statistical significance was assessed using two-tailed t test. * indicates P < 0.05, ** indicates P < 0.01, and n.s. indicates not significant. (C) The chromatin was prepared from HEK293 cells and the ChIP was performed using mouse IgG or PRMT5 antibody or rabbit IgG or FAM47E antibody. The association of PRMT5 and FAM47E with the promoters of PRMT5 target genes was investigated by analyzing the immunoprecipitated DNA using quantitative RT-PCR. Data are presented as fold enrichment relative to the control IgG binding and the values represent the mean of three independent experiments, with error bars representing standard deviations.