Effects of manipulation of several intracellular signaling pathways on basal and FSH-induced ERK2 phosphorylation in cultured granulosa cells of prehierarchical (6–8 mm) follicles.

<p>Top panel: Representative western blot analysis of phosphorylated and total ERK2 in granulosa cells treated without or with 10 ng/ml FSH in combination with PKA activator or inhibitor (10 μM Forskolin or 20 μM H89, respectively), or PKC activator or inhibitor (20 nM PMA or 10 μM GF109203X, respectively), or PI3K inhibitor (20 μM LY294002) or mTOR inhibitor (10 μM Rapamycin), or AMPK activator (1 mM AICAR). β-actin was used as loading control. Bottom panel: Quantitative analysis of relative protein abundance of phosphorylated ERK2 by densitometry using Image Lab software (Version 4.1, Bio-Rad laboratories). Relative protein abundance was normalized to total ERK2. Data are expressed as fold differences ± SEM of three independent experiments using tissues from different hens and are compared to control cells. ^*, <i>P</i> < 0.05 compared to control cells; ^#, <i>P</i> < 0.05 compared to cells only stimulated by 10 ng/ml FSH.