Fig. 3

Insulin’s Suppressive Role on hESC Decidualization Operates Through the PI3K/AKT Pathway. (A-B) For a period of 2 (A) and 4 (B) days, human endometrial stromal cells (hESCs) were treated with 0.5 mmol/L db-cAMP to induce decidualization, both with and without insulin. The phosphorylation status of endometrial AKT (Ser473) was determined by Western blot analysis. (C-D) The presence or absence of the PI3K inhibitor LY294002 was factored in during the 2 (C) and 4 (D) day-decidualization period, after which the protein levels of IGFBP1 were evaluated using Western blot. Densitometric quantification of protein expression was conducted with ImageJ software. The summarized results are expressed as mean ± SD for a sample size of n = 3, with * P < 0.05 indicating statistical significance. (E) Over a 4 day interval, hESCs underwent differentiation with 0.5 mmol/L db-cAMP and 200 nmol/L insulin, with the inclusion or exclusion of 5µmol/L LY294002. mRNA expression levels were quantified by qRT-PCR. These results are also shown as mean ± SD for n = 3 samples, with * P < 0.05 representing statistically significant differences