Figure 4.

siRNA p5 Functions in a Transcription-Silencing Manner

Hep 3B or Hep G2 cells were transfected with 50 nM siRNAs and co-incubated with DMSO or either (A and B) 40 nM trichostatin A (TSA) or (C and E) 7.5 μM 5′ azacytidine. (A and C) Hep 3B and (B) Hep G2 cells were harvested 72 h later and assessed for LDLR expression by qRT-PCR. (D) Hep 3B cells were transfected with 100 nM p5 and processed for ChIP with H3K27me3 or its mouse IgG control. DNA elutes were amplified with primers toward the target region of p5 or with an off-target primer set, located 8 kb upstream of the LDLR promoter in the LDLR gene. (E) Hep 3B treated with 7.5 μM 5′ azacytidine were harvested 72 h later and assessed for BM450697 expression by qRT-PCR. Data are the mean ± SD of three (A and B) or two (C–E) independent experiments with triplicate treated conditions. Two-way ANOVA (A–C) or one-way ANOVA (D) with the post hoc Tukey’s test was performed. *p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001.