Research Paper Volume 12, Issue 2 pp 1304—1321

Figure 2. CCND1 is a direct target of AURKB. (A) Quantitative real-time PCR analysis of the effect of AURKB knockdown by siRNA on the mRNA levels of CCND1, CDC16, CDC6, CDC26, CCNB2, CCNF, p27 and E2F1 in SGC7901 and BGC823 cells relative to those in the negative control (NC) cells. The results shown are the means ± SDs of three independent experiments; **, P < 0.01 compared with the negative control. (B) Western blot analysis showing the effect of AURKB knockdown by siRNA on the expression of CCND1 in SGC7901 and BGC823 cells. HSP70 was the loading control. (C) Western blot analysis showing the effect of AURKB overexpression on the expression of CCND1 in SGC7901 and BGC823 cells. HSP70 was the loading control. (D) Quantitative real-time PCR analysis of the effect of AURKB overexpression on the mRNA levels of CCND1 in SGC7901 and BGC823 cells. The results shown are the means ± SDs of three independent experiments; **, P < 0.01 compared with the negative control. (E–F) Chromatin immunoprecipitation assays showing the effect of AURKB knockdown on H3S10ph (E) H3R8me2s, H3K9me2, or H3K9me3 (F) enrichment in the CCND1 promoter in SGC7901 and BGC823 cells. Normalized inputs of SGC7901 and BGC823 chromatin DNA were pulled down by antibodies against H3S10ph or negative immunoglobulin G (IgG). The results shown are the means ± SDs of three independent experiments; **, P < 0.01 compared with the negative control.