G5045

BZA was acquired from Sigma (B7273, purity > 98%). Phenylephrine (PE, P1240000) was obtained from Sigma-Aldrich. Anti-PPAR-α (sc-9000) and anti-PCNA (sc-7907) were purchased from Santa Cruz Biotechnology. The following first antibodies were purchased from Cell Signaling Technology: anti-AKT (#4691), anti-phospho-AKT (#4060), anti-GSK3β (#9315), anti-phospho-GSK3β (#9323P), anti-ERK (#4695), anti-phospho-ERK (#4370P), anti-P38 (#9212P), anti-phospho-P38 (#4511P), anti-AMPKα (#2603P), and anti-phospho-AMPKα (#2535). Anti-GAPDH (#ab8245), anticalcineurin (CaN) (#ab90540), and anti-NFAT1 (#ab2722) were obtained from ABCAM. Anti-α-actinin was acquired from Millipore. The secondary antibodies were purchased from LI-COR Biosciences. The PPAR-α antagonist (GW6471, G5045), PPAR-β/δ antagonist (GSK0660, G5797), and PPAR-γ antagonist (GW9662, M6191) were all purchased from Sigma-Aldrich. All other chemicals were of analytical grade.

Human prostate carcinoma DU145 (ATCC; HTB-81), PC3 cells (ECCAC; HTB-81) and their drug-resistant sub-lineages were routinely cultivated in DMEM/F12 HAM (Sigma, St. Louis, MO) medium supplemented with 10% fetal bovine serum (FBS) and antibiotics [19 (link),38 (link)]. Human umbilical vein endothelial cells (HUVEC; Life Technologies Corporation, Carlsband, CA, USA) were cultured (up to six passages) in endothelial basal medium (EBM; Lonza, Basel, Switzerland) supplemented with 10% fetal bovine serum (FBS) and supplement cocktail (hydrocortisone, recombinant hEGF, bovine brain extract, gentamicin, amphotericin-B; all from Lonza [58 (link)]). For endpoint experiments, media supplemented with DCX and/or FF were added to cancer cell cultures at the concentrations given in the text. Culture media were supplemented with: docetaxel (DCX; 0.125–50 nM), mitoxantrone (MTX; 62.5–2000 nM) and/or fenofibrate (FF; 5–25 μM; F6020, Sigma, Saint Louis, MO, USA), GW6471 (10 μM; G5045), N-acetyl-L-cysteine (NAC; 1 mM; A9165, Sigma), elacridar (100 nM); sulfinpyranoze (500 μM), Fumitremorgin C (10 μM), 3-methyladenin (3MA; 0.5 μM; all from Sigma) at the time points indicated in the text. Chemical inhibitors were administrated at the time points indicated in the text and at the concentrations that secure their specific action and the lack of cytotoxic effects.