Enhanced chemiluminescence ecl

Cinobufagin was obtained from the National Institute for the Control of Pharmaceutical and Biological Products (Beijing, China) and dissolved in Dulbecco's modified Eagle's medium (DMEM) (Gibco, Grand Island, NY, USA) to each working dose. Fetal bovine serum (FBS) was purchased from Hangzhou Sijiqing Biological Engineering Material Co., Ltd. (Hangzhou, Zhejiang, China). Chloroquine (CQ), 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), N-acetyl cysteine (NAC) and 3-methyladenine (3-MA) were purchased from Sigma-Aldrich (St. Louis, MO, USA). The Annexin V-FITC Apoptosis Detection Kit I was obtained from BD Biosciences (San Diego, CA, USA). Hoechst 33258 was purchased from Promega Corp. (Madison, Wi, USA). Lipofectamine 2000 transfection reagent was purchased from Invitrogen Life Technologies (Carlsbad, CA, USA). Antibodies against caspase-3, caspase-8, caspase-9, cleaved PARP, phospho-JNK, total JNK, phospho-p38, total p38, LC3B, p62, GAPDH, enhanced chemiluminescence (ECL) and western blotting kits were purchased from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA).

Dulbecco's modified Eagle's medium (DMEM), fetal bovine serum (FBS), and antibiotics (penicillin and streptomycin) were purchased from Gibco BRL (Rockville, MD, USA). Anti-β-actin, 3-(4,5-dimethylthiazol-2yl)-2,5-diphenyltetrazolium bromide (MTT) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Anti-CTGF, anti-JNK, anti-pJNK, anti-p38, anti-pp38, anti-ERK1/2, anti-pERK1/2, goat anti-rabbit IgG, goat anti-mouse IgG, donkey anti-goat IgG, enhanced chemiluminescence (ECL), Egr-1, and control siRNA were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). AGE-BSA was obtained from Calbiochem (Darmastadt, Germany), PD98059, U0126, SP600125 and SB203580 were purchased from Calbiochem (San Diego, CA, USA). Lipofectamine2000TM was purchased from Invitrogen (Carlsbad, CA, USA). Recombinant CTGF was purchased from R&D system (Minneapolis, ME, USA) and α-tubulin from Sigma (St. Louis, MO).