Research Paper Volume 12, Issue 11 pp 10578—10593

Figure 2. CARM1 is a positive regulator of CCNE2 gene in NSCLC cells. (A) ChIP analysis of human CCNE2 promoter by antibodies against CARM1, H3R17me2a, H3R26me2a or IgG in NC or CARM1-silenced PC9 and HCC827 cells. Relative enrichment of CARM1, H3R17me2a and H3R26me2a marks on the promoter regions was analyzed by real-time PCR assays. The data were presented as means ± SDs of three independent experiments; **P < 0.01, ^#P > 0.05. (B) The luciferase activity of CCNE2 promoter reporter was significantly increased when CARM1 (100 ng, 200 ng, 500 ng and 1000 ng) was transfected into PC9 and HCC827 cells. The CCNE2 promoter reporter luciferase activity was normalized to beta-galactosidase activity. The data were shown as means ± SDs of three independent experiments; **P < 0.01. (C) The mRNA levels of CCNE2 was downregulated in CARM1-depleted PC9 and HCC827 cells by Real-time PCR assays. β-actin was used as an internal control. The data were presented as means ± SDs of three independent experiments; **P < 0.01. (D) The protein levels of CCNE2 was downregulated in CARM1-depleted PC9 and HCC827 cells by Western blot. GAPDH was used as loading control.