Research Paper Volume 11, Issue 10 pp 3094—3116

Tyrosine nitrations impaired intracellular trafficking of FSHR to the cell surface and FSH-induced Akt-FoxO3a signaling in human granulosa cells

Figure 5. FoxO3a inactivation was required for FSH-mediated cell survival. KGN cells transfected with or without NC siRNA or FSHR siRNA were incubated with or without FSH (1 nM, 4 hrs). (A) The apoptotic indices were measured using in Situ cell death detection kit. (B) The caspase-3 enzyme activity was assayed using a commercial kit. Relative gene expressions of FasL (C) and Bim (D) were determined by RT-PCR (β-actin as internal standard). (E) Relative protein expressions of p-Akt and Akt were determined by immunoblots (β-actin as internal standard). On the other hand, KGN cells transfected with or without FoxO3a-WT and FoxO3a-S253A were incubated with or without FSH (1 nM, 4 hrs). (F) The apoptotic indices were measured using in Situ cell death detection kit. (G) The caspase-3 enzyme activity was assayed using a commercial kit. Relative gene expressions of FasL (H) and Bim (I) were determined by RT-PCR (β-actin as internal standard). Open triangle: p<0.05 vs. Ctrl; Bold triangle: p<0.05 vs. FSH (n = 3–6).