Methylation of the Tumor Suppressor Protein, BRCA1, Influences Its Transcriptional Cofactor Function
Figure 4
Methylation status of BRCA1 alters BRCA1-DNA interactions at specific promoters in vivo.
(a) MDA-MB-231 cells were treated with AdOx (30 µM) in order to observe BRCA1 methylation inhibition upon treatment. Two milligram of MDA-MB-231 whole cell protein extract was immunoprecipitated with anti-BRCA1 and anti-IgG antibodies, separated on a 4–20% gel by SDS-PAGE, and probed with anti-K methyl antibody. Blot was stripped and reprobed with anti-BRCA1 antibody. Input represents 1/10 of immunoprecipitated material. Results are representative of two independent experiments. (b) MDA-MB-231 cells were treated with AdOx (30 µM) for 48 hours prior to being collected for ChIP analysis. Antibodies used for ChIP were anti-BRCA1 (10 µg), anti-IgG (10 µg), and anti-histone H3-phosphorylated at S10 (H3-pS10, 5 µg). PCR products were run on a 2% agarose gel and visualized with ethidium bromide staining. Results are representative of two independent experiments.