Figure 7
SM decreased decidualization of human eSCs. (A) Human eSCs were incubated either under 1 g or SM for 1 day, after which they were induced to differentiate in the presence of 0.5 mM 8-Br-cAMP under 1 g for 1 day. Cells were lysed and subjected to a quantitative real time PCR (qRT-PCR) analysis. (B) The cells were treated as in (A), induced to decidualization for 4 days, (C) stained using senescence associated β-galactosidase staining kit, and photographed under microscope. Scale bar = 50 μm. (D) The cells were treated with 8-Br-cAMP under 1 g or SM for 1 day, lysed, and analyzed by qRT-PCR. (E) The cells were differentiated in the presence of 8-Br-cAMP for one day and shifted to either 1 g or SM for one day in the presence 8-Br-cAMP. The cells were then subjected to qRT-PCR. (F) The cells were differentiated as in (D) and shifted to either 1 g or SM for one day in the absence of 8-Br-cAMP. (G) The cells were incubated under SM with or without 5 μg/ml SC-79 for 1 day, after which they were induced to differentiate in the presence of 8-Br-cAMP under 1 g for 1 day. The cells were lysed and subjected to a qRT-PCR analysis. Abbreviations: simulated microgravity (SM); simulated microgravity for 24 h before the induction of differentiation (Pre-SM); prolactin (PRL); insulin-like growth factor binding protein 1 (IGFBP1). Data are expressed as mean ± SD, with paired t-tests performed as indicated. °P < 0.05 versus undifferentiated control; *P < 0.05 versus differentiated control without Pre-SM; •P < 0.05 versus differentiated cells with Pre-SM.
