Mitochondrial membrane potential detection

Bortezomib (Velcade^®) and antibodies against Bax, tubulin, cytochrome c, GAPDH, caspase-3, caspase-9, and PARP were purchased from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA). Antibodies against BID and cleaved caspase-3 were purchased from Cell Signaling Technologies (Beverly, MA, USA). XIAP, cIAP1, and caspase-8 antibodies were purchased from R and D (USA). BD Cytofix/Cytoperm plus fixation and permeabilization solution kit with BD GolgiPlug, propidium iodide (PI) staining solution, annexin V binding buffer, mitochondrial membrane potential detection (JC-1) kit, stain buffer (FBS), annexin V-FITC antibody, H2AX (pS139)-Alexa Fluor 647 antibody, rabbit anti-active caspase-3- Bv605 antibody and PARP cleaved form-AF700 antibody were obtained from BD Biosciences (NJ, USA). Apoptotic DNA-ladder kit was obtained from Roche (Penzberg, Germany).

The HTB-132 MCF-7 and MCF-12 cell lines were obtained from ATCC^® (Manassas, VA, USA); LIVE/DEAD^TM BacLightTM Viability Kit, and annexin V, Alexa Fluor^TM 488 conjugate was purchased from Invitrogen (Eugene, Oregon); Mitochondrial Membrane Potential Detection was obtained from BD Biosciences (Torreyana Rd., San Diego, CA 92121, USA). The DMEM culture medium was obtained from Sigma-Aldrich (St. Louis, MO, USA). Bovine fetal serum was purchased in Gibco (Waltham, MA, USA). The Fmoc-amino acids, Rink amide resin, dicyclohexylcarbodiimide and 1-hydroxy-6-chlorobenzotriazole were purchased from AAPPTec (Louisville, KY, USA). Acetonitrile, trifluoroacetic acid, dichloromethane, diisopropylethylamine, N, N-dimethylformamide, ethanedithiol, isopropanol, methanol, triisopropylsilane were purchased from Merck (Darmstadt, Germany). SPE SupelcleanTM columns were purchased from Sigma-Aldrich (St. Louis, MO, USA) and Silicycle^® SiliaPrepTM C18 columns were kindly donated by EcoChem Especialidades Químicas (Waterloo, QC, Canada).