Fig. 1: mTORC1 and mTORC2 differentially regulate 8-Br-cAMP-induced decidualization.

a Human endometrial stromal (hES) cells were induced to differentiate for 4 days in the presence of 0.5 mM 8-Br-cAMP. On day 0, 2, or 4 of differentiation, the cells were lysed and subjected to western blotting. b, c On day 0 or 2 of differentiation, the cells were lysed, subjected to immunoprecipitation against raptor (b) or rictor (c) and were analyzed by western blotting. d–g hES cells were infected with lentiviruses expressing two different raptor shRNAs (d, e), rictor shRNAs (f, g), or scrambled shRNA. They were then selected by puromycin for 5 days and differentiated for 2 days in the presence of 0.5 mM 8-Br-cAMP. d, f The cells were lysed and analyzed by either quantitative RT-PCR or western blotting. Human GAPDH was used to normalize gene expression. e, g Cells were photographed under bright-field illumination. Round-shaped cells were scored as differentiated cells. Scale bar = 50 μm. a–g Data are representative of three to four independent experiments. d-g Data are expressed as the mean ± SD, with paired t-tests performed as indicated. *P < 0.05, **P < 0.01