Fig. 2

Effects of pretreatments with AKT-siRNA-, LY294402-, and rapamycin-pretreatment on in vitro osteogenic/dentinogenic differentiation of SCAP. SCAP were pretreated with or without AKT-siRNA (20 nM) (a–d), LY294402 (50 μM) (e–h), and rapamycin (100 nM) (i–l) for 3 days and were cultured under osteogenic/dentinogenic condition. a, e, i Representative western blot images of the expression of AKT and p-AKT (a, e) and mTOR and p-mTOR (i) in SCAP 1 week after the osteogenic/dentinogenic induction. b, f, j Representative western blot images of the expression of RUNX2, BGLAP, and DSPP in SCAP 1 week after the osteogenic/dentinogenic induction. c, d, g, h, k, l Alizarin Red-S staining shows the calcium deposition in SCAP 4 weeks after osteogenic/dentinogenic induction. Representative images of Alizarin Red-S staining. Bars = 25 mm (c, g, k). Calcium accumulation assay. n = 5 for all groups. *P < 0.05. Graph bars show the means ± SEM (d, h, l)