Hdac4

Antibodies used were against MEF2D (BD Bioscience), H3K27ac (ab4729; Abcam, Cambridge, MA, USA). H3K27me3 (ab195477, Abcam) HDAC4 (Paroni et al., 2004), γH2AX (9718, Cell Signalling, Leiden, The Netherlands), TP53 (DO‐7; Dako, Santa Clara, CA, USA), LT SV40 (sc‐147, Santa Cruz), Lamin B1 (ab16048, Abcam), p21 (CP74, Sigma), RACK1 (sc‐17754, Santa Cruz, Dallas, TX, USA), GFP (Paroni et al., 2004).

The phalloidin, JC-1 and DCFH-DA probes were bought from Life Technology (St. Louis, MO, USA). PX-12, DAPI and isoprenaline hydrochloride (Iso) were purchased from Sigma-Aldrich (USA). Rhodamine-conjugated secondary antibodies were obtained from Jackson ImmunoResearch Laboratories (West Grove, PA, USA). STVNa was obtained from Chemical Development Laboratories of Key Biological Pharmaceutical Company (Dongguan, China). Trizol was obtained from Generay (Shanghai, China). Antibodies against Drp1, Trx1, Prdx2 and GAPDH were acquired from Cell Signaling Technology (Beverly, MA, USA). HDAC4 was obtained from Abcam (Cambridge, MA, UK). The Fis1 antibody and HRP-conjugated secondary antibodies were acquired from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

For Western blot analysis, brain tissues and cells were lysed in RIPA lysis buffer with phosphorylase inhibitor and protease inhibitor for 15 minutes on ice. After centrifugation at 14000 rpm for 15 minutes, the supernatant was collected and the protein content of the samples was determined by the Bradford method. Equal amounts of protein were loaded onto 10% SDS-PAGE gels and blotted onto PVDF membranes. Membranes were blocked with 4% nonfat milk and incubated with primary antibodies against HDAC4 (1 : 1000; Abcam, Cambridge, MA, USA), HDAC4 phosphorylated at Ser-632 (1 : 200; Santa Cruz Biotechnology, Heidelberg, Germany), HIF-α (1 : 200; Abcam, Cambridge, MA, USA), and VEGFa (1 : 200; Santa Cruz Biotechnology, Heidelberg, Germany) overnight. β-Actin (1 : 4000; Abcam, Cambridge, MA, USA) was used as a loading control. After washing three times with Tris-buffered saline, the membranes were incubated with HRP-conjugated anti-rabbit or anti-mouse secondary antibodies (1 : 2000) for 2 hours at room temperature. Specific binding was detected with enhanced chemiluminescence reagents. The blots were analyzed with ImageJ analysis software, and P-HDAC4 was normalized to total HDAC4 for comparisons.

The following primary antibodies were used: proliferating cell nuclear antigen (PCNA) was purchased from Cell Signaling Technology; cyclin A was purchased from Santa Cruz Biotechnology; HDAC4, ATM2C1, Ki67, cyclin A2, H3K27me3, and MAP2 were purchased from Abcam; and LB509 and AT8 were purchased from ThermoFisher Scientific. PHF1 was a generous gift from Dr. Peter Davies (Albert Einstein College of Medicine, Bronx, NY).
Secondary antisera conjugated with fluorescent Alexa Fluor dyes 488 and 647, and Cy3 were purchased from ThermoFisher Scientific and Jackson ImmunoResearch.

PCNA, p38, phospho-p38 (Thr180/Tyr182), ERK, phospho-ERK (Thr202/Tyr204), JNK, phospho-JNK (Thr183/Tyr185), NFκB p105/50, NFκB p65, phospho-NFκB p65 (Ser536), RelB, Akt, phospho-Akt (Ser473), and phospho-Akt (Thr308) antisera were purchased from Cell Signaling Technology (Danvers, MA, USA); CD45, Iba-1, GFAP, γ-H2AX, HDAC4, MAP2, GAPDH, 8-oxoguanine, and Ki67 antisera were purchased from Abcam (Cambridge, MA, USA); and cyclin A1 and c-Rel antisera were purchased from Santa Cruz Biotechnology (Dallas, Texas, USA). Secondary antisera conjugated with fluorescent Alexa dye 488 and 647 and Cy3 were purchased from Life Technologies and Jackson ImmunoResearch (West Grove, PA, USA). HRP-conjugated secondary antibodies were purchased from Cell Signaling Technology and Life Technologies.