Fig. 6. Acute effect of Kras^G12D expression in cultured granulosa cells. (A) Expression of Kras^G12D mRNA in LSL-Kras^G12D granulosa cells after infection with Ad-CMV-Cre. (B) Levels of phospho-ERK1/2 and phospho-AKT post-infection. (C) LSL-Kras^G12D granulosa cells infected with Ad-CMV-Cre and control vectors (Ad-CMV-GFP) for 48 hours were stimulated with FSH, forskolin (Fo) or amphiregulin (AR) for 20 minutes. Each agonist induced ERK1/2 phosphorylation in control granulosa cells, but the responses were reduced in the cells expressing KRAS^G12D.