Anti caspase 3

Mouse kidney cells (TCMK-1) were a gift from Dr. T.J. Lee (Yeungnam University, Korea). Primary culture of human mesangial cells (Cryo NHMC) were purchased from Clonetics (San Diego, CA, USA), and other cell lines are from the ATCC (Manassas, VA, USA). All cells were maintained in Dulbecco’s modified Eagle’s medium, and supplemented with 10% FBS, 5% antibiotic solution, and 100 μg/mL gentamycin. Garcinol and lactacystin was purchased from Enzo Life Sciences (Ann Arbor, MI, USA). Recombinant human TRAIL and z-VAD-fmk were purchased from R&D (Minneapolis, MN, USA). Calbiochem supplied N-acetyl-l-cysteine (NAC), trolox, and MG132 (San Diego, CA, USA). The following antibodies were used: anti-XIAP, -Bcl-2, -Mcl-1, -survivin, -Cbl, and -Itch (Santa Cruz Biotechnology, Santa Cruz, CA, USA); anti-c-FLIP (ALEXIS Corporation, San Diego, CA, USA); anti-PARP, -DR5, -PSMA5, -PSMD4/S5a, -cleaved caspase-3 (Cell Signaling Technology, Beverly, MA, USA); anti-caspase-3 (Enzo Life Sciences, Ann Arbor, MI, USA); anti-DR4 (Abcam, Cambridge, MA, USA); anti-actin (Sigma-Aldrich, St. Louis, MO, USA). Santa Cruz Biotechnology (Santa Cruz, CA, USA) and Bioneer (Daejeon, Korea) supplied DR5 siRNA and GFP siRNA (control siRNA), respectively. Sigma Chemical Co. supplied other reagents (St. Louis, MO, USA).

Antibodies used include anti-Flag M2 (Sigma, F3165), anti-TOM20 (Abcam, ab78547), anti-BAG5 (Cusabio, CSB-PA890743HA01HU), anti-Actin (Sigma, A2066), anti-GAPDH (Cell Signaling Technology, 14C10), anti-GFP (Abcam, Ab290), anti-Parkin (Cell Signaling Technology, 2132S), anti-Caspase-3 (Enzo, ADI-AAP-113-D), anti-Cleaved Caspase-3 (Cell Signaling Technology, 9664S), anti-Tubulin (Cell Signaling Technology, 2148S), anti-Mcl-1 (Cell Signaling Technology, 4572S), anti-HA-tag antibody conjugated sepharose beads (Cell Signaling Technology 3956), and anti-HA (Sigma, 11867423001).

EDHB was purchased from Sigma-Aldrich (St. Louis, MO) and dissolved in ethanol for biochemical assays. Anti-AKR1C antibodies were obtained from Santa Cruz Biotechnology (Santa Cruz, CA). Anti-caspase-3 and anti-caspase-9 antibodies were procured from Enzo Life Sciences (NY). Anti-LC3A/B and anti-Beclin antibodies were purchased from Cell Signaling Technology (Boston, MA). Anti-β-actin antibodies were obtained from Sigma-Aldrich (St. Louis, MO). Anti-BNIP3 and anti-NDRG1 antibodies were purchased from Abcam (Abcam, MA).

Artichoke capsules (Aboca, Sansepolcro, Italy); Pemetrexed (ALIMTA, Eli Lilly and Company, Indiana, USA) and Cisplatin (Pfizer Pharmaceuticals Group, New York, USA) were dissolved according to the manufacturer's instructions. The following primary antibodies were used: anti-cleaved PARP (Asp214) (Cell Signaling, # 9541); anti-caspase 7 (Cell Signaling, #9492); anti-caspase 3 (Enzo life Science, #31A1067); anti-beta actin (A-2228, SIGMA); β-catenin antibody (clone CAT-5H10, # 18–0226. Zymed). Secondary horseradish peroxidase-conjugated was purchased from Santa Cruz; secondary antibody for immunofluorescence Alexa Fluor 594 (mouse) conjugated was obtained from Molecular Probes (Inc, Eugene, OR, USA). ECL reagent (Amersham, GE Healthcare, Piscataway, NJ, USA) was employed for the chemo-luminescence detection. Annexin V FITC (0, 2 μg/ml) (Abcam, ab-63556); DAPI staining (Sigma) was used for nuclear detection.

The EDHB chemical compound was purchased from Sigma-Aldrich (St. Louis, MO) and dissolved in ethanol for biochemical assays. Anti-NDRG1 and anti-BINP3 antibodies were purchased from Abcam (Abcam, MA). Anti-caspase-3 and anti-caspase-9 antibodies were purchased from Enzo Life Sciences (Life Sciences, NY). Anti-HIFα, anti-Beclin, anti-ATG12 and anti-LC3 antibodies were purchased from Cell Signaling Technologies (Danver, MA). Anti-AKR1C1, anti-Bcl-2, anti-p53, anti-TOM20 and anti-p21 antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA).