Figure 3.

GDF9 and BMP15 regulate SFRP4 via SMAD3. (A) Primary human cumulus cell (CCs) were treated with the SMAD2/3 inhibitor SB431542 (1µM) for 1 h before the addition of GDF9 and BMP15 (GB: 5 ng/ml each) and FSH (50 ng/ml). Cells were harvested 48 h later. (B) Cells were treated with the SMAD3 inhibitor SIS3 (S3: 5 µM), the SMAD1/5/8 inhibitor LDN-193189 (S1/5/8, 100 nM) or SB431542 (S2/3, 1µM) for 1 h before the addition of GDF9 plus BMP15 (GB: 5 ng/ml each). For both graphs, SFRP4 mRNA levels were determined by qPCR and expressed relative to Rpl19. Columns represent the mean ± SEM, columns with different letters differ significantly by one-way ANOVA analysis with Bonferroni correction, a–b P < 0.05, a–c, b–c P < 0.01 (n = 8).