Figure 3.

SHH-Induced Promoting Effects on Endometrial Stem Cells Are Mediated through the FAK/ERK1/2 and/or PI3K/Akt Signaling Pathways

A schematic showing an overview of the molecular mechanisms of SHH-mediated effects on endometrial stem cells (A). Endometrial stem cells were stimulated for 10 min with or without SHH (4 μM) treatment. The endometrial stem cells were then lysed, and the protein expression was analyzed by western blotting using antibodies targeting the phosphorylated forms of FAK, ERK1/2, PI3K, and Akt. The phosphorylation levels of these signaling molecules were significantly increased in cells treated with SHH (B and C). Differentially expressed genes from nonproliferative cells and proliferative cells (GEO: GSE62564 and GSE85047) were analyzed using Ingenuity Pathway Analysis (IPA) software (https://www.ingenuity.com/) to predict the activation state (either activated or inhibited) of the Akt (D) and ERK1/2 (E) signaling pathways. The GEO database (https://www.ncbi.nlm.nih.gov/geo/) was analyzed to further verify the significant correlation between stemness and the activation state of Akt and ERK1/2 signaling pathways (F–H). β-Actin was used as the internal control. The data represent the mean ± SEM from three independent experiments.