Figure 4.

Loss of IRS2 reduces PI3K/AKT and MAPK activation in HESC during decidualization. Following siRNA mediated knockdown of IRS2 and 24-hour stimulation with DC, protein expression was investigated by Western blot (A) and immunofluorescence (B). (A) Whole cell lysates were analyzed by Western blot and probed with antibodies specific for phosphorylated and total ERK1/2 and phosphorylated and total AKT. Calnexin was used as a loading control. Densitometry analysis is provided (right). (B) Immunofluorescent staining using antibodies specific for phosphorylated ERK1/2 and phosphorylated AKT. DAPI was used as a nuclear stain. Images provided are 40× magnification. (C) Gene expression analysis was performed with RNA isolated from HESC treated with DC and DMSO or either 10μM U0126 or 100nM MK2206 for 72 hours using primers specific for IGFBP1, WNT4, and PRL. 36B4 was used for normalization. Data are represented as the mean fold induction ± SEM. *P < 0.05 relative to DC+DMSO.