Pan acetyl h3

Following the drug treatments, cells were collected and resuspended in RIPA lysis buffer (Beyotime, China) and lysed by ultrasonic instrument. Then equal protein amount of the cell extracts was analyzed on a 12% SDS-polyacrylamide gels and transferred onto PVDF membranes. Membranes were blotted with primary antibodies targeting the following proteins: LSD1 (Sangon Biotech, Shanghai, China), pan-acetyl-H3 (Millipore, USA), Mono-Methyl-Histone H3 (Lys4), Di-Methyl-Histone H3 (Lys4), Tri-Methyl-Histone H3 (Lys4), Mono-Methyl-Histone H3 (Lys9), Di-Methyl-Histone H3 (Lys9), Tri-Methyl-Histone H3 (Lys9), PARP [poly(ADP-ribose) polymerase], Caspases-3, P21, P27, P57, Bcl-xL, Bcl-2, Bax and LC3B (all from Cell Signaling Technology, USA). β-actin (Abcam, USA) and Histone H3 (Abcam, USA) were used as internal controls. Then, membranes were incubated in HRP-conjugated secondary antibody and visualized using the HRP Substrate (Millipore, USA) by Image Quant LAS-4010 system (GE Healthcare, USA).

Harvested cells were resuspended in a lysis buffer containing 50 mM Tris/HCl (pH 7.5), 150 mM NaCl, 1% Triton X-100, 1% sodium deoxycholate, 0.1% SDS and 0.1 mM PMSF. Proteins were analyzed by SDS-PAGE and Western blotting. Membranes were incubated with primary antibodies against pan-acetyl-H3 (Millipore, USA), PARP [poly(ADP-ribose) polymerase], γ-H2AX (Ser139), caspases-3, Bcl-xL, Bcl-2, Bax, H3, β-actin (all from Cell Signaling Technology, USA). Blots were visualized using the ECL (Millipore, USA) with Image Quant LAS-4010 (GE Healthcare, USA).