BRCA1 methylation status alters protein-protein interactions at the 504-802 region.

<p>(<b>a</b>) Schematic of BRCA1 504-802 primary sequence depicting important protein-protein interactions and domains that could be affected by the methylation of this region. (<b>b</b>) 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 or anti-IgG antibodies, separated on a 4-20% gel by SDS-PAGE, and western blotted using antibodies against Sp1 and BRCA1 proteins. Input represents 1/10 of immunoprecipitated material. Results are representative of two independent experiments. (<b>c</b>) MDA-MB-231 cells were treated with AdOx (30 µM) and whole cell extract separated on a 4-20% gel by SDS-PAGE, and probed with anti-Sp1 antibody. Densitometry was averaged from three independent immunoblots.</p

Decreased levels of PRMT1 alters BRCA1 promoter binding <i>in vivo.</i>

<p>(<b>a</b>) HeLa cells were transfected with different concentrations of PRMT1 siRNA (10, 25, 50 nM) following manufacturer's instructions. Results are representative of two independent experiments. (<b>b</b>) HeLa cells transfected with 50 nM Luc or PRMT1 siRNA were collected for ChIP analysis. Anti-BRCA1 (10 µg), anti-IgG (10 µg), and anti-histone H3-phosphorylated at S10 (H3-pS10, 5 µg) antibodies were used for ChIP analysis. PCR products were run on a 2% agarose gel and visualized with ethidium bromide staining. Results are representative of two independent experiments.</p