Figure 2.

Dose-dependent effects and time course of the effects of gonadotropin-releasing hormone agonist (GnRHa) on 3β-hydroxysteroid dehydrogenase (3β-HSD) expression. Rat Leydig cells were treated with GnRHa at various concentrations for 24 h or at a concentration of 100 nmol l⁻¹ for various time courses. GnRHa was replaced along with the medium every 12 h. Cells were harvested for total RNA or protein extractions, and 3β-HSD expression was detected by real-time PCR (a, b) and Western blot analysis (c, d). The quantity of 3β-HSD mRNA was normalized as a ratio to the amount of Rp119 mRNA. The intensities of the protein bands in the Western blots were quantified, and 3β-HSD protein levels were normalized to that of β-actin. Data are presented as mean±s.e.m. of three separate experiments. * P < 0.05 compared with the control; #, P < 0.05 compared with 0 h.