Fig. 8.

Androgen regulation of CYP4F2 expression in LNCaP cells. a, b LNCaP cells were grown in steroid-deprived media for 8, 12, or 24 h in the presence of vehicle (control); in DHT, 10 nM; or in complete media (FBS 10%). Then, they were collected and subjected to RNA and protein extraction for the quantitation of CYP4F2 mRNA normalized to HPRT-1 (qPCR) (a) and the analysis of CYP4F2 protein expression normalized to β-tubulin (Western blot) (b). The expression levels were calculated as the values relative to that of a calibrator sample (control, 8 or 12 h for qPCR and WB, respectively). c Cells were grown in steroid-deprived media for 24 h, in the presence of vehicle or DHT, 10 nM, for the measurement of intracellular 20-HETE levels by ELISA. d, e LNCaP cells were grown in complete media for 24 h in the presence of vehicle (control) or enzalutamide, 10 μM, and subjected to the analysis of CYP4F2 mRNA expression normalized to HPRT-1 (d) and protein expression normalized to β-tubulin (e). a n = 2–3, in duplicates (ANOVA p = 0.0005; **p < 0.01 vs. all conditions except DHT, 12 h). b n = 3 (ANOVA p = 0.0023; *p < 0.05 vs. time-related control). c n = 2, in duplicates (t test p = 0.0297). d n = 2, in duplicates (t test p = 0.045). e n = 2 (t test p = 0.0193). Values are presented as the mean ± SEM