Figure 7
PGE2 activates AKT in an Epac-dependent manner. (A) hBMSCs were cultured continuously in the absence (−PGE2) or presence (+PGE2) of PGE2 (10 nM), and AKT phosphorylation levels determined by Western blot analysis at day 7, 8 and 9. *p < 0.01 as compared to −PGE2 using Student’s t-test. (B) hBMSCs were cultured continuously in the absence (−PGE2) or presence (+PGE2) of PGE2 (10 nM) with or without Epac inhibitor ESI-09 (10 μM) or PKA inhibitor PKI (10 μM), and AKT phosphorylation levels determined by Western blot analysis at day 9. *p < 0.01, **p < 0.001 as compared to -PGE2 using ANOVA. # p < 0.01, ## p < 0.001 using ANOVA. In both cases, GAPDH served as a loading control and representative cropped blots shown. (C) hBMSCs were cultured continuously in the absence (−PGE2) or presence (+PGE2) of PGE2 (10 nM) with or without Epac inhibitor ESI-09 (10 μM), and RUNX2, ALP, BGLAP and MGP expression levels determined by RT-qPCR at day 17. *p < 0.05, **p < 0.001 as compared to -PGE2 using ANOVA. # p < 0.01, ## p < 0.001 using ANOVA. The data represent triplicate determinations and were replicated at least two times. All values are presented as mean ± S.D.
