Protein samples were diluted in SDS sample buffer and resolved by PAGE. 50 µg total protein was transferred to PVDF (Biorad) using 10 mM CAPS, pH 10.5, 15% methanol, 3.5 mM DTT transfer solution for 17 hours at 20 mA and 20 V limits. Membranes were blotted with affinity purified anti-Bptf at 1∶10000 dilution [14], M2 anti Flag monoclonal antibody at 1∶5000 (Sigma), anti Smad2 and Smad2-phos (Cell Signaling Technologies) 1∶1000, anti SNF2H/L (Abcam) 1∶5000, anti-CBP and p300 1∶100 (Abcam) overnight at 4°C then anti rabbit or mouse HRP (Amersham) at 1∶20000 dilution for 2 hrs at RT in PBS with 5% NFDM and 0.1% tween 20 through out.
