Fig. 3: The inhibitory effects of GnRH on PI3K/Akt signaling in vitro and in vivo.

Schematic representation of the experimental protocol as described in the materials and methods section (a). Endometrial stem cells were treated for 10 min with or without GnRH (1 µM). The cells were then lysed, and the protein contents were analyzed by western blotting using antibodies targeting the phosphorylated forms of PI3K and Akt. The phosphorylation levels of these signaling molecules were significantly decreased in stem cells treated with GnRH (b). Endometrial stem cells were treated with 1 µM GnRH alone or in combination with shRNA targeting GnRH-R; subsequent changes in the phosphorylation levels of PI3K and Akt were measured via western blotting (c). Mice were treated daily for 21 days with GnRH (0.1 mg/kg, subcutaneously) or vehicle (PBS). Stem cells were isolated from mouse adipose tissues, and changes in the phosphorylation levels of PI3K and Akt were measured via western blotting (d). β-actin was used as the internal control. The data are presented as the mean ± SD of three independent experiments