Prostate cancer PC-3 and 22RV1 cell lines, and benign prostate BPH1 line were obtained from ATCC (Manassas, VA) as described.16 (link),17 (link) C4–2 cell line was obtained from UroCor Inc (Oklahoma City, OK).18 (link) All cell lines were tested for authentication at Genetica DNA Lab (Burlington, NC) and were used within passages of 15–25. Cells were cultured in RPMI 1640 media containing 10% fetal bovine serum and 1% penicillin/streptomycin in a 5% CO2-humidified atmosphere at 37°C. Biotin labeling of Alternol was carried out in house. Succinate assay kit and enzymatic function assay kits for PDHC, KGDHC, FH, and MDH2 were obtained from BioVision (Milpitas, CA). Docetaxel was obtained from Cayman Chemical (Ann Arbor, MI). Alternol was obtained from Sungen Bioscience as reported14 (link) (Shantou, China).
Antibodies for Caspase-3, DLAT, DLST, FH, MDH2, GAPDH, HSPD1, HYOU1, HSP90AB1, HSPA8, ILF2, NPM1, PDIA6, ATP5A1, ANXA2, TUBA1B, and LAMR1 were obtained from Cell Signaling Technology (Danvers, MA). TUNEL immunostaining kit, ETC complex proteins antibodies and Ki-67 antibodies were obtained from Abcam (Cambridge, MA). Beta-actin antibody was obtained from Sigma (St. Louis, MO). Streptavidin-agarose beads and secondary antibodies were obtained from Santa Cruz Biotech (Santa Cruz, CA).
Antibodies for Caspase-3, DLAT, DLST, FH, MDH2, GAPDH, HSPD1, HYOU1, HSP90AB1, HSPA8, ILF2, NPM1, PDIA6, ATP5A1, ANXA2, TUBA1B, and LAMR1 were obtained from Cell Signaling Technology (Danvers, MA). TUNEL immunostaining kit, ETC complex proteins antibodies and Ki-67 antibodies were obtained from Abcam (Cambridge, MA). Beta-actin antibody was obtained from Sigma (St. Louis, MO). Streptavidin-agarose beads and secondary antibodies were obtained from Santa Cruz Biotech (Santa Cruz, CA).