Fig. 2

Involvement of PKA, PKC, MEK/ERK and PI3K pathways in LH-induced expression of DCN. MGCs collected during IVF procedures were cultured for 4 days with daily medium exchanges. Then, MGCs were either stimulated with (a) vehicle (control), hCG 10 U/ml, 10 μM forskolin (FSK), 20 nM PMA, or with a combination of 10 μM FSK and 20 nM PMA for 24 h. Cells were pretreated for 1 h with H-89 (10 μM), U0126 (10 μM), or LY 294002 (10 μM), and then stimulated with 10 U/ml hCG, 10 μM FSK, or 20 nM PMA for 24 h. Total mRNA was subjected to qPCR with DCN (a-d) primers and b-actin (housekeeping gene) primers. DCN was quantified by qPCR and normalized to β-actin expression. Data represent the mean ± SEM of three independent experiments. Different letters represent significant differences (p < 0.05). e Schematic representation of LH/hCG signaling cascade that mediates DCN expression in MGCs