Figure 5

Effects of MEK inhibitor on CYP17A1 mRNA expression and androstenedione production in bovine theca cells. Subconfluent cultures were pretreated with MEK inhibitor (U0126, 10 μM) for 30 min. Then they were stimulated with LH (100 ng/ml) for 12-24 h. Control cells (CTL) were cultured in the absence of added treatments. RT-PCR was conducted using CYP17A1 and 36B4 (internal control) primers using total RNA isolated from the cells. The mRNA level of CYP17A1 were expressed as ratio to 36B4 values (Top). Culture media were also assayed for androstenedione by EIA (Bottom). Data are the mean ± SEM (n = 4). Each experiment was reproduced at least three times. Different letters represent statistically significant differences of means (P < 0.05).