Bcl xl s

Total protein was extracted from cells lysates using RIPA lysis buffer and loading buffer as previous described.17 Proteins were separated with SDS‐PAGE and incubated with specific antibodies. Protein bands were visualized and quantified with an Odyssey system (Pierce, Waltham, MA, USA). The antibodies used were: AKT, phospho‐AKT (Ser473), phospho‐p70S6K1 (Thr389), ERK, phospho‐ERK (Thr202/Tyr204), MEK1/2, phospho‐MEK1/2 (Ser271/221), IκBα, phospho‐IκBα (Ser32/36), STAT3, phospho‐STAT3 (Tyr705), caspase 9, caspase 8, and PARP, all obtained from Cell Signaling Technology (Danvers, MA, USA). BCL‐xL/S, MCL‐1, NF‐κB(p65), NF‐κB(RelB), and CXCR4(4G10) were bought from Santa Cruz Biotechnology (Dallas, TX, USA); Ac‐H3K9 obtained from Active Motif (Carlsbad, CA, USA); Phospho‐CXCR4 (S339) (ab74012) was purchased from Abcam (Cambridge, UK); β‐actin was obtained from Millipore (Burlington, MA, USA); The BCL‐2 antibody was purchased from Dako (Agilent Technologies, Santa Clara, CA, USA). Fluorescent‐conjugated secondary anti‐rabbit or anti‐mouse antibodies were purchased from Enzo life sciences.

4-Hydoxy-2-nonenal was bought from Cayman Chemical Co. (Ann Arbor, MI). N-Acetylcysteine was purchased from Sigma Chemical Co. (St. Louis, MO). DMEM/F12, DMEM and fetal bovine serum (FBS) were obtained from GIBCO BRL (Grand Island, NY). DCFH-DA was purchased from Beyotime Institute of Biotechnology (Haimen, China). Antibodies against PARP, MKP-1, Nrf2, p53, Bax, Bcl-xL/S, GAPDH, histone H3 and β-actin were obtained from Santa Cruz Biotechnology (Santa Cruz, CA). Antibodies against caspase-3, cleaved-caspase-3, total and phospho (p)-ERK1/2 (Thr202/Tyr204), JNK, p-JNK, p38 MAPK, p-p38 MAPK, eIF2α, p-eIF2α (Ser51), LC3A/B and PP2A were products of Cell Signaling Technology (Beverly, MA). Peroxidase-conjugated goat anti-rabbit and goat anti-mouse secondary antibodies were purchased from Huaxingbio Biotechnology Co. (Beijing, China). The annexin V-FITC&PI kit was from Jiamay Biotechnology (Beijing, China). Kinases inhibitors, including ERK1/2 inhibitor U0126, JNK inhibitor SP600125, p38 MAPK inhibitor SB202190, and protein degradation inhibitor MG132 were obtained from Cell Signaling Technology (Beverly, MA). PERK inhibitor GSK2606414 was from Selleck Chemicals (Houston, TX). Other reagents used in this study were ordered from Sigma, unless otherwise stated.