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. 2001 Apr 17;98(9):4990–4995. doi: 10.1073/pnas.101617298

Figure 3.

Figure 3

Expression of p16 and p14 and ChIP analysis of MBD2 and acetylated histones H3 and H4 in colon cancer cell lines and HeLa. (A) p16 transcripts were coamplified by RT-PCR, with the GAPDH transcripts as an internal control; 1 μg of total RNA from HCT116, SW48, HCT15, and HeLa cells was used. PCR products (5 μl) were separated by electrophoresis on a 2% agarose gel, and positions of the bands for p16 (250 bp) and GAPDH (308 bp) are indicated on the left. (B) Expression of p14 was analyzed by RT-PCR after coamplification of p14 transcript (235 bp) with GAPDH (308 bp). (C) PCR analysis of DNA in chromatin immunoprecipitated with anti-MBD2 and anti-acetylated histone H3 and H4 for p16 (395 bp), p14 (508 bp), and Alu (362 bp). Experiments were performed on the following cell lines: HCT116 with one allele methylated (M+) for p16 and p14 M−; SW48 with p16 M+ and p14 M−; HCT15 with p16 M+, p14 M+, and HeLa cells with p16 and p14 M−. (D) Statistical analysis (mean ± SD of at least three experiments) of the enrichment of the bound DNA fraction compared with the unbound DNA fraction for p16 and p14, for each antibody and in each cell line.