A. BRMS1 co-immunoprecipitates HDAC3. FLAG-HDAC3 was co-transfected into COS-7 cells with either pCMV-myc (V) or pCMV-myc-BRMS1(B) using Lipofectamine 2000 (Invitrogen). The cell lysate was immunoprecipitated with 1 μg anti-FLAG Ab (Sigma). The precipitated proteins were resolved by SDS-PAGE and immunoblotted with 1:500 dilution anti-myc Ab (BD Clontech). Independently the cell lysate was immunoblotted with anti-FLAG antibody to confirm expression of FLAG-HDAC3. B. BRMS1 reduces acetylation of p65. MDA-MB-435 cells were transfected with pCMV-myc (V) or pCMV-myc-BRMS1 (B) and immunoprecipitated with anti-p65 antibody. The immunoprecipitate was resolved on SDS-PAGE, transferred to PVDF membrane and immunoblotted with anti-acetyl lysine antibody. The lysate shows that the levels of p65 are not altered by BRMS1.