Figure 2.

IRS2 is required for HESC decidualization. (A) Gene expression analysis was performed following IRS2 knockdown and 72-hour stimulation with DC using primers specific for IGFBP1, BMP2, WNT4, HAND2, and PRL. 36B4 was used for normalization. Data are represented as the mean fold induction ± SEM. *P < 0.05 relative to siCtrl. (B) Following knockdown and stimulation with DC for 72h, IGFBP1, BMP2, WNT4, and HAND2 expression was determined by Western blot. Calnexin is provided as a loading control. Densitometry analysis is provided (below). (C) IRS2 knockdown was performed and cells were stimulated for 6 days with DC and stained with phalloidin to bind actin filaments. DAPI was used as a nuclear stain. Images provided are 40× magnification. (D) IRS2 knockdown was performed and cells were exposed for 24 hours of proliferative (E+P) and decidualization (DC; E+P+cAMP) conditions. Immunofluorescent staining of Ki67 was performed and DAPI was used as a nuclear stain. Ki67 positive cells were counted using ImageJ and expressed as a percentage of total cells. Representative panels are shown; 20× magnification.