Hdac3

The A2780 cell line was purchased from American Type Culture Collection (ATCC), and SAHA was purchased from Selleckchem (Houston, TX, USA). Primary antibodies against Ac-H2A, Ac-H2B, Ac-H3, Ac-H4, HDAC2, HDAC3, HDAC4, DNMT3A, PRMT1, SUV39H1, MDMX, p53, p21^WAF1/CIP1, p27^Kip1, AURKB, CDC25C, GADD45A (1:1000) and Alexa Fluor 488 dye were purchased from Cell Signaling Technology (Danvers, MA, USA). The MDM2 antibody (1:500) was purchased from Santa Cruz Biotechnology (Dallas, TX, USA). β-actin (1:5000), secondary antibodies (anti-rabbit, anti-mouse), horseradish peroxidase (HRP) conjugate, and dimethyl sulfoxide (DMSO) were purchased from Sigma Aldrich (St. Louis, MO, USA). Nitrocellulose membrane (NC) (0.45 µm) was purchased from Amersham (GE Healthcare Life Sciences, Marlborough, MA, USA) and KPL LumiGlo Reserve chemiluminescent substrate was purchased from SeraCare Life Sciences (Milford, MA, USA).

After antigen obtainment, the 4 μm-thick slices were incubated with antibodies against Nrf2 (1:200; Cell Signaling Technology), NeuN (1:100; Boster Biotech, Wuhan, China), LC3 (1:100; Cell Signaling Technology) and HDAC3 (1:100; Cell Signaling Technology) overnight at 4°C. Following washing, they were then incubated with secondary antibodies for an additional 1 h at room temperature. The cell nuclei were stained with 4’,6-diamidino-2-phenylindole (DAPI). A pathologist who is blind to experiments randomly selected five ROI under a high magnification optical microscope (400×; Leica) to observe the positive staining cells surrounding injury areas. Five random ROIs were selected for quantification and the mean was used for the statistical analysis.

LRS chambers were obtained from consented regular blood donors at Children’s Hospital Colorado, Aurora, Colorado, USA. PBMCs were lysed and protein complexes were purified by centrifugation. Protein complexes were incubated with rec-Protein G-sepharose beads (Invitrogen) and antibodies directed against ETV6 (Sigma Prestige) overnight at 4°C on a rocking platform. After incubation, beads were sequentially washed 3 times with 1× PBS (Gibco). Beads were boiled for 10 minutes in 2× SDS-PAGE sample buffer (Bio-Rad) to elute pulldown products for Western blot analysis. Protein eluate was run on Tris-Glycine SDS-page gel (Bio-Rad) and transferred to nitrocellulose membranes. Membranes were blocked in tris-buffered saline with 0.1% Tween-20 containing 5% milk at 4°C for 1 hour and were probed with HDAC3 (Cell Signaling Technologies), NCOR2 (Abcam), and α-tubulin (Abcam) primary antibodies on a shaker at 4°C overnight, followed by 1 hour of room temperature incubation with HRP-conjugated secondary antibodies (Invitrogen). Chemoluminescence was recorded using the automated Gel Doc XR+ system (Bio-Rad).

SNOH-3, with purity >98%, was synthesized in Medicine Chemistry Laboratory at Shenyang Pharmaceutical University (see Figure 3a and Supplementary data). SAHA were obtained from Sigma (St. Louis, MO, USA). These agents were dissolved in DMSO to 100 mM and stored at −20 °C. Before treatment, the stock solution is diluted to different concentrations. The final concentration of DMSO in cultures is 0.1% (v/v) or less. MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) and propidium idodide (PI) were purchased from Sigma. The primary antibodies against HDAC1, HDAC3, HDAC6, HDAC8, Histone 3, Histone 4, p21^WAF1/CIP1, PARP, Caspase-3, Survivin, XIAP, KLF4, VEGF, and β-actin were got from Cell Signaling Technology. The primary antibodies against CD31 were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA). The primary antibodies against Ac-H3 and Ac-H4 were purchased from Millipore (Boston, MA, USA). The HDAC1 Silencer Select Validated siRNA was got from Life Technologies (Waltham, MA, USA).

p-Akt (Ser473) (#4060), p-Akt (Thr308) (#13038), Akt (#4691), Cyclin D1 (#2978), p-p38 (Thr180/Tyr182) (#4511), p-ERK1/2 (Thr202/Tyr204) (#4370), ERK1/2 (#9102), p38 (#8690), Bcl-2 (#3498), K17 (#4543), p-IKKα/β (Ser176/180) (#2697), IKKβ (#8943), p-IκBα (Ser32) (#2859), p-p65 (Ser536) (#3033), p65 (#8242), p-p70S6K (#9234), p70S6K (#9202), HDAC1 (#5356), HDAC2 (#5113), HDAC3 (#3949), HDAC4 (#7628), HDAC6(#7558), H3 (#4499), p-SAPK/JNK (Thr183/Tyr185) (#4668), and SAPK/JNK (#9252) antibodies were purchased from Cell Signaling Technologies, USA. mTOR (# sc-1549), ICAM-1 (#sc-8439), β-Actin (#sc-47778), COX-2 (#sc-1745), PCNA (#sc-7907), Ki-67 (#sc-15402), p-STAT3 (#sc-8059), STAT3 (#sc-8019), anti-Goat (#Sc-2354), anti-mouse (#Sc-2061), and anti-mouse (#Sc-2030) IgG-horseradish peroxidase (HRP) antibodies were procured from Santa Cruz Biotechnology, USA.