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. 2010 Oct 29;5(10):e13732. doi: 10.1371/journal.pone.0013732

Figure 5. Effect of downregulation of Cbx7 on expression of p16/Arf/p15, cell growth and p16 methylation.

Figure 5

(A), analysis of CBX7 expression in PC3 cells by Western blot assay 72 hours after CBX7 knockdown. Protein extracted from 293T cells was used as P16 positive control (P16-p.c.). (B), the growth curves of PC3 cells stably transfected with the scramble control shRNA vector (shR-Ctrl) or shRNA-Cbx7 vector (shR-Cbx7) in MTT assay. (C), analysis of transcription of Cbx7, Arf, and p15 by quantitative RT-PCR. The statistical significance of the differences of mRNA relative copy number between cells stably transfected with shR-Ctrl and shR-Cbx7 were labeled above the top of one of the columns. (D), analysis of p16 methylation by MSP in the Cbx7 stably knockdown PC3 cells. Genomic DNA from AGS and MGC803 cell lines was used as positive control of methylated and unmethylated p16 respectively. (E), comparison of H3K9me3 level by ChIP assays within various locations as illustrated in Figure 3A.