Anti hdac

Lysates, cytosolic and nuclear extracts were electrophoresed on a 4-12% polyacrylamide gel (Invitrogen) and proteins were transferred to an Immobilon membrane (Millipore). Membranes were blotted with anti-IκBα (Cell Signaling, #4814), anti-IκBβ (R&D systems, AF5225), anti-p50 (Abcam, ab7971), anti-p65 (Cell Signaling, #8242), anti-IL-1α (R&D systems, AF-400-NA), anti-HDAC (for nuclear extracts, Cell Signaling, #5356), anti-Calnexin (for lysate, Enzo Life Sciences ADI-SPA-860), and anti-GAPDH (for cytosolic extracts, Cell Signaling, #5174). Densitometric analysis was performed using Image Studio (LiCor).

TAX (purity ≥ 98%), 5-azadeoxycytidine (5-aza, purity ≥ 97%), trichostatin A (TSA, purity ≥ 98%), bacteriological agar, Eagle’s basal medium (BME) and TPA were obtained from Sigma-Aldrich (St. Louis, MO, USA). Fetal bovine serum (FBS), Dulbecco’s modification of eagle’s medium (DMEM), minimum essential medium (MEM) and trypsin-EDTA solution were obtained from Gibco Laboratories (Gaithersburg, MD, USA). The primary antibodies anti-Nrf2, anti-HO-1, anti-NQO-1 and anti-β-actin were acquired from Santa Cruz Biotechnology (Santa Cruz, CA, USA). The primary antibodies anti-DNMT (DNMT1, DNMT3a and DNMT3b) were obtained from IMGENEX (San Diego, CA, USA). The primary antibodies anti-HDAC (HDAC1, HDAC2, HDAC3, HDAC4, HDAC5, HDAC6, HDAC7 and HDAC8) were obtained from Cell Signaling Technology (Danvers, MA, USA).