Figure 2.

Effect of hormones on the steroid profile of H295R cells. H295R cells are usually grown in DMEM/Ham’s F-12 medium enriched with NU-I serum (according to ATCC guidelines). To remove hormones we stripped NU-I serum by charcoal treatment (see Methods). H295R cells were then grown in normal growth medium with charcoal-stripped NU-I serum medium and the steroid profile was assessed after 48 hours. Steroid production was labeled with [³H] pregnenolone for 90 min and extracted steroids were resolved by TLC. A representative TLC is shown on the left and the quantitative analysis of steroids showing no change is shown on the right. Data are the mean ± SEM of two independent experiments. Prog, progesterone; Δ4A, androstenedione; Preg, pregnenolone; 17OH Preg, 17α-hydroxypregnenolone; 17OH Prog, 17OHP; 11DOC, 11-deoxycortisol.