Fig. 4. Kras^G12D downregulates genes essential for granulosa cell differentiation and ovulation. (A) qRT-PCR of ovulation-related genes from mouse whole ovary mRNAs. Six ovaries from different animals were analyzed. (B) Kras^G12D downregulated the expression of Fshr and prevented the FSH-induced expression of Lhcgr in cultured granulosa cells. Expression of Kras^G12D was induced by infecting the cells with an adenoviral vector encoding Cre recombinase (Ad-Cre). FSH (100 ng/ml) was added to the medium of cells infected, or not, with Ad-Cre. NT, non-treated. Ppib was amplified by RT-PCR in the same samples, as loading control.