Anti active caspase 3

Anti-active caspase-3, -MMP-9, -MMP-13, -TNF-α and TNF-α were purchased from R&D Systems, Inc., (Heidelberg, Germany). Antibodies to phospho-specific p65 (NF-κB)/(Ser536) were purchased from Cell Technology (Beverly, MA, USA). Antibodies to collagen type II and alkaline phosphatase-linked sheep anti-mouse and sheep anti-rabbit secondary antibodies for immunoblotting were purchased from Millipore (Schwalbach, Germany). Anti-TNF-β was obtained from eBioscience (Frankfurt, Germany). Anti-Ki67 and secondary antibodies used for fluorescence labelling were purchased from Dianova (Hamburg, Germany). Anti-cyclooxygenase-2 was purchased from Cayman Chemical (Ann Arbor, MI, USA). Monoclonal anti-β-Actin was obtained from Sigma-Aldrich (Munich, Germany). Sox9 antibody was purchased from Acris Antibodies GmbH (Hiddenhausen, Germany). Polyclonal antibody against Sirt1 was purchased from Abcam PLC (Cambridge, UK).TNF-β and polyclonal rabbit Anti-TNF-β were obtained as described [40 (link)]. Bacteria-derived recombinant human TNF-α and LT-α (TNF-β), both purified to homogeneity with a specific activity of 50 million units/mg, were kindly gift from Genentech, Inc. (South San Francisco, CA).

Sections were de-paraffinized and heated in citrate buffer for improved antigen retrieval and further incubated with anti-islet-1/2 (40.2D6) 1:50; anti-gag (AMV3C2, 1:200; both from Developmental Studies Hybridoma Bank, University of Iowa), anti-BrdU (Sigma; 1:1000); anti-active-caspase-3 (R&D; 1:500) and visualized using biotinylated secondary antibodies (donkey-anti-mouse; -anti-rabbit; -anti-rat; 1:100; Dianova, Hamburg, Germany) and diaminobenzidine (Vectastain Peroxidase ABC-kit 6100, Vector Laboratories, CA, United States).

Anti-phospho-DNA-PKcs and -histone H4 antibodies were from Abcam (Cambridge, MA, UN). Anti-DNA-PKcs antibody was the product of BBI Life Science (Shanghai, China). Anti-IKKβ antibody was from Merck Millipore (Billerica, MA, USA). Anti-Akt1 and -XIAP antibodies were purchased from Cell Signaling Technology (Danvers, MA, USA). Anti-RIP1 and -PARP antibodies were obtained from BD Bioscience (San Diego, CA, USA). Anti-active caspase 3 and -cIAP1 antibodies were the products of R&D (Minneapolis, MN, USA). Cisplatin, puromycin, butylated hydroxyanisole (BHA), N-acetyl-L-cysteine (NAC), and Chloroquine (CQ) were purchased from Sigma-Aldrich (St. Louis, MO, USA). The pan-caspase inhibitor zVAD and DNA-PK inhibitor NU7026 were the products of Merck Millipore (Billerica, MA, USA). Small interfering RNA (siRNA) targeting different proteins and negative control siRNA were from Guangzhou RiboBio Co. (Guangzhou, China). siRNA transfection reagent was from Polyplus transfection (Illkirch, France). TRIzol reagent was from Invitrogen (Carlsbad, CA, USA).