Fig. 5

Insulin’s Inhibition of hESC Decidualization is Governed by the PI3K/AKT-NR4A Pathway. Human endometrial stromal cells (hESCs) were prompted to undergo differentiation for 2 days (A) and 4 days (B) with 0.5 mmol/L dibutyryl-cAMP (db-cAMP) and varying concentrations of insulin (0, 10, 50, 200 nmol/L). This was followed by a qRT-PCR analysis to evaluate the expression of NR4As. (C) Corresponds to the cell treatment detailed in (A), after which the cells were lysed for Western blot analysis. These results are displayed as mean ± SD for n = 3 samples, where # P < 0.05 indicates statistical significance compared to the undifferentiated control (Ctrl), and * P < 0.05 is in relation to the differentiated control (De). In a continuation of the experiment, hESCs were induced to differentiate for 2 (D) and 4 (E) days with 0.5mmol/L db-cAMP alongside 200nmol/L insulin, with and without the addition of 5µmol/L LY294002. mRNA expression was quantified by qRT-PCR and NR4A1 protein levels were determined via Western blot (F). These findings are expressed as mean ± SD for n = 3 samples, with * P < 0.05 denoting statistical significance