Figure 3

Leptin induced MAPK pathway in human granulosa cells. (A) The expression of leptin receptor in human immortalized granulosa cells were monitored by semi-quantitative RT-PCR. Upper bands showed the expression of short form leptin receptor (Ob-Ra) in 4 different passages of immortalized granulosa cells. Lower band is to detect the expression of long form leptin receptor (Ob-Rb) in immortalized granulosa cells. GAPDH was used as internal standard. Cells were treated with leptin (1000 ng/ml) for different time scale (5, 15, 30, 60 min) and cell lysates were collected and subjected to Western blot analysis to monitor the expression of phosphorylated ERK1/2 (B), phosphorylated p38 (C) and phosphorylated JNK (D). Total ERK1/2, p38, JNK and β-actin were detected as internal standard. Upper bands are Western blotting representative datas from three independent experiments. Lower bars were integrated optical density (IOD) of target phosphorylated protein expression with target total protein normalization. Each data point in the figure represents the mean ± SD of three independent experiments. * above the bars indicates that those groups differ significantly from untreated control (P < 0.05). (ctrl = control).