Fig. 5. Estrogen/GPER/Gαi signaling increases HIF-1α expression by altering protein stability of HIF-1α.

A WPMY-1 were pretreated with 10 mg/ml cycloheximide (CHX) and then treated with vehicle control or 1 nM G1 for 0–12 h. HIF-1α protein levels were analyzed by western blot. The right graph is the quantified HIF-1α degradation curve. B Plvthm/shGPER-WPMY-1 were treated with or without MG132 (5 mM) for 3 h in the presence of 1 nM G1, and then HIF-1α protein levels were analyzed by western blot. C WPMY-1 were treated with 1 nM E2 or 1 nM G1 with or without 0.5 µM MK2206 for 72 h, and the expression of the indicated genes was detected by western blot. GAPDH or β-actin was used as the loading control.