Figure 5
Effect of rh adiponectin on phosphorylation of MAPK ERK 1/2 (A) and AMPK (B) in bovine granulosa cells. Granulosa cells from small follicles were cultured for 24 h in enriched McCoy's 5A medium supplemented with 10% FBS, starved of serum for 18 h and then incubated in serum-free medium with or without adiponectin (10 μg/mL) from 1 to 120 min. 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 or anti-phospho-AMPKα. The blots were stripped and reprobed with antibodies against ERK2 or AMPKα, respectively. Bands on the blots were quantified and the phosphorylated/total protein ratio was calculated. Values represent means ± SE relative to the basal state from at least three independent experiments. Bars with different superscripts are significantly different (P < 0.05).