FIG. 4.

Quantitative real-time PCR analysis of the effect of MEHP (200μM) on mRNA levels of a selected set of genes involved in Leydig cell cholesterol metabolism and steroidogenesis, of LH-R (Lhcgr) and Insl3 genes in E13.5 (white bars) and E18.5 (black bars) testes after 3 days of culture in the absence (A) or presence (B) of 100 ng/ml oLH. The results were calculated by the delta-delta Ct method using an external standard (luciferase) added during RNA extraction as an endogenous reference. The levels of mRNA are expressed as mean of relative unit ± SEM of six to nine cultures with the control animals having a value of 1, with each sample processed in duplicate. *p < 0.05, **p < 0.01, ***p < 0.001 versus corresponding control value in the paired Student's t-test.