Figure 5

EGF-induced granulosa cell proliferation and activation of downstream signaling are modulated by the expression of HOXA7. KGN cells were transiently transfected with scrambled siRNA or HOXA7 siRNA. SVOG cells were transiently transfected with control vector or HOXA7 plasmid. KGN cells (A) and SVOG cells (C) were reseeded into a 96-well plate after 36 h of transfection. Cells were serum starved for 12 h and incubated in serum-free medium containing 100 ng/mL EGF and/or AG1478, or were left untreated (Control) for 48 h. Cell viability was measured by the MTT assay. KGN cells (B) and SVOG cells (D) were serum starved for 12 h after 36 h transfection and incubated in serum free medium containing 100 ng/mL EGF and/or AG1478, or left untreated (Control) for 10 min. The phosphorylation of ERK1/2 or Akt was determined by Western blotting with specific antibodies. The data derived from at least three separate sets of experiments were standardized to the corresponding control, and the statistical results are presented in the column graphs. a, P < 0.05 compared with control; b, P < 0.05 compared with control siRNA or control vector transfection and EGF treatment. c, P < 0.05 compared with EGF treatment alone.