Chemicals and Reagents
Dibutyryl cAMP (dbcAMP) and 22-hydroxycholesterol were obtained from Sigma-Aldrich. Pregnenolone was obtained from Steraloids (purity 99.8%). Testosterone antibody was obtained from Fitzgerald Industries International. Bovine LH (USDA-bLH-B-6) was provided by the United States Department of Agriculture Animal Hormone Program [16].
Polyclonal rabbit anti-cytochrome P450scc cholesterol side-chain cleavage enzyme (P450scc) antibody was obtained from Millipore (#ABS235). Polyclonal rabbit anti-steroidogenic acute regulatory protein (StAR) antibody was obtained from Santa Cruz Biotechnology (#25806). Monoclonal mouse anti-gp91phox antibody was obtained from BD Transduction Laboratories (#611414). Polyclonal rabbit anti-4-hydroxy-2-nonenal (HNE) antibody was obtained from Alpha Diagnostic International (#HNE-11S). Polyclonal rabbit anti-glutathione peroxidase-1 (GPx-1) antibody was obtained from Abcam (#22604). Monoclonal mouse anti β-actin antibody was obtained from Sigma Chemical (#A5316). The horseradish peroxidase-conjugated anti-mouse (#NA931V) or anti-rabbit (#NA934V) donkey second antibodies were obtained from GE Healthcare Biosciences.