Fig. 3: FSH-mediated suppressive effects in endometrial stem cells are regulated through Akt and/or ERK1/2 signaling cascades.

A schematic summary of the roles of FAK/ERK1/2 and/or PI3K/Akt signaling cascades in regulating the FSH-mediated inhibitory effects on endometrial stem cells is described (a). Cells were treated with or without FSH at 30 IU/ml for 10 min. Treated endometrial stem cells were washed with PBS and then lysed. Subsequent changes in the phosphorylation levels of Akt, PI3K, FAK, and ERK1/2 were measured by western blotting (b, c). Endometrial stem cells were treated with FSH (30 IU/ml) alone or were concomitantly transfected with an shRNA specifically targeting FSHR. Subsequent changes in the phosphorylation levels of PI3K, Akt FAK, and ERK1/2 were measured by western blotting (d, e). Differentially activated genes in rapidly proliferating cells and nonproliferating cells were analyzed using IPA software to investigate the activation status (intermediate, inactivate, or activate) of AKT1 (GSE62564) (f) or MAPK1/3 (ERK1/3) (GSE21034, GSE44752) (g)-associated signaling molecules/transcription factors. Furthermore, the GEO data repository was used to assess the relationship between various FSH-enhancing conditions and the expression levels of AKT or MAPK1/3 (h). β-Actin was used as the internal control. All experiments were performed in triplicate. Data are presented as the mean ± standard deviation (SD). *p < 0.05; **p < 0.005; and ***p < 0.001 (two-sample t test).