Figure 3

Time- (A) and dose- (B) dependent effects of GnRH (Leuprolide acetate) on the rate of Smad3 activation (phospho-Smad3; pSmad3). HES and ESC were incubated under serum-free condition and treated with GnRH (0.1 μM) for 5, 15 and 30 min, or with GnRH at 0.01 to 10 μM for 15 min. The cell lysates were prepared and analyzed by immunoblotting for pSmad2/3 and β-actin as loading control, shown from a representative experiment. Bar graphs show the mean ± SEM of fold change in the rate of Smad3 activation from three independent experiments. In Fig. A: denotes b, c, d, b', c', and d' and in Fig. B, c, d, e, b', c', d,' and e'are significantly different from a and a', respectively (p < 0.05).