Figure 8.
Effect of rh adiponectin on progesterone secretion by bovine COC (A) and phosphorylation of MAPK ERK1/2 in these complexes (B). Progesterone secretion (A) was measured by RIA protocol in culture medium of COC (20 COC/200 μL medium/well) after 24 h of IVM in TCM 199 serum-free medium supplemented or not with rh adiponectin (10 μg/mL) +/- insulin (10-8 M). The data are expressed as the amount of progesterone (ng/mL) secreted per 24 h. The results, expressed as means ± SE, are representative of four independent cultures. (B) IVM of COC (15 COC/well) was performed in TCM 199 serum-free medium supplemented or not with rh adiponectin (10 μg/mL) ± insulin (10-8 M) for 1 h. Protein extracts were separated by electrophoresis on 12% (w:v) SDS-polyacrylamide gel. After transfer to nitrocellulose membranes, the proteins were probed with anti-phospho-ERK1/2. The blots were stripped and reprobed with antibodies against vinculin. Bands on the blots were quantified and the phospho-ERK1/2/vinculin ratio was calculated. Values represent means ± SE relative to the basal state from three independent experiments. Bars with different superscripts are significantly different (P < 0.05).