Anti gcn5

Antibodies of this study were as follows: anti-HBXIP (Abcam, UK), anti-HOXB13 (Abcam), anti-β-actin (Sigma-Aldrich, USA), anti-Flag-tag (Sigma-Aldrich), anti-STAT3 (ImmunoWay Biotechnology Company, USA), anti-GFP (Sigma-Aldrich), anti-GCN5 (Proteintech, USA), anti-acetylated lysine (Aviva Systems Biology, CA), anti-p300 (Santa Cruz Biotechnology, USA), anti-ER-α (ImmunoWay Biotechnology Company), anti-HSC70 (Proteintech), anti-Ki67 (Santa Cruz Biotechnology), and IL-6 neutralizing antibody (Abcam). Trichostatin A (TSA) and cycloheximide (CHX) were separately purchased from Beyotime Biotechnology (China) and MedChem Express (USA). 4OH-TAM and ASA were purchased from Sigma-Aldrich.

For immunofluorescence studies of HDAC1 and GCN5, cells were treated with the psychostimulants and opioid with or without piracetam and grown in chamber slides. For immunostaining cells were fixed with 4% paraformaldehyde, followed by permeabilization with 0.2% Triton X-100 in PBS for 15 min at room temperature and then blocked with 5% normal goat serum at 4°C for 1h and then incubated with antibodies: anti-HDAC1 (Cell Signaling Technology) and anti-GCN5 (Proteintech) overnight at 4°C. After washing with 1X PBS, cells were incubated with anti-mouse-IgG Alexa Fluor® 633 for GCN5 and anti-rabbit-IgG Alexa Fluor® 546 for HDAC1 for 2h. The cellular nuclei were stained with 4’6-diamidino-2-phenylindole (DAPI). Slides were examined with a Nikon C1plus confocal microscope.