Figure 4

Effects of PKA inhibitor, MEK inhibitor and PI3K inhibitors on Akt phosphorylation in bovine theca cells. Subconfluent cultures were pretreated with PKA inhibitor (H89, 3 μM), MEK inhibitor (U0126, 10 μM), or PI3K inhibitors (wortmannin, 0.1 μM; LY294002, 25 μM) for 30 min. Then they were stimulated with LH (100 ng/ml) for 24 h. Control cells (CTL) were cultured in the absence of added treatments. Cell lysates (20 μg) were subjected to SDS-PAGE and Western blot using anti-phosphorylated-Akt antibody (Phospho-Akt) or anti-total-Akt antibody (Total-Akt). Representative images (Top) and densitometric data of phospho-Akt contents (Bottom), expressed as a ratio of phospho-Akt to total-Akt, are shown. Values show the mean ± SEM for three experiments. Each experiment was reproduced at least three times. Different letters denote significant differences of means (P < 0.05).