Rabbit anti acetyl h3

Adherent and non-adherent cells were collected from drug-treated dishes and total protein extracted by Tris-SDS lysis buffer, quantified by Lowry and used for immunoblotting. Antibodies used in the study were rabbit anti-Bcl-xL (1:1000, #2764; Cell Signaling), mouse anti-Bcl-2 (1:500, Dako; #M0887), mouse anti-PCNA (1:2000,#MS106-PO; Neomarker), rabbit anti-p89-PARP (1:1000, #9541; Cell Signaling) and rabbit-anti-Acetyl-H3 (1:1000, #06-599; Millipore). Equal protein loading between lanes was confirmed by staining membranes with Ponceau S prior to immunoblotting.

Zebrafish embryos were homogenized in RIPA buffer with protease and phosphatase inhibitors (Beyotime Institute of Biotechnology, Shanghai, China). The supernatants were collected by centrifuging at 12000 rpm at 4 °C for 15 min. The supernatants were mixed with loading buffer, and equal amounts of protein were loaded onto 15% SDS-PAGE gels. When the separated proteins were transferred to the PVDF membranes, the PVDFs were incubated with primary antibodies and then with an HRP-conjugated goat anti-rabbit IgG antibody (1:5000, AQ132P; Sigma-Aldrich). The primary antibodies were used as follows: rabbit anti-H3 (1:1000, #4499; Cell Signaling Technology), rabbit anti-acetyl-H3 (1:1000, #06-599; Millipore), rabbit anti-H3K27me3 (1:1000, #9733; Cell Signaling Technology), rabbit anti-H3K9me3 (1:1000, # 13969; Cell Signaling Technology), rabbit anti-GFP (1:1000, #2956; Cell Signaling Technology), and rabbit anti-β-actin (1:2000, SAB2100037; Sigma-Aldrich). Protein bands were analysed by densitometry using Quantity One.