Fig. 6

RNAs from hSHED-EVs regulated hSHED-EV-enhanced TRET-associated hBMMSC properties and functions. hBMMSCs were incubated with RNase-pretreated and MOCK-pretreated SHED-EVs (RNase-EV and MOCK-EV, respectively). a Expression of TERT in hBMMSCs by RT-qPCR. b Telomerase activity in hBMMSCs assayed by RQ-TRAP. H.I., heat-inactivated group. c Expression of CD146 and percentages of BrdU⁺ cells in hBMMSCs after FCM and BrdU incorporate assays. d Representative images of mineralized nodules after Alizarin Red staining 4 weeks after osteogenesis. Bars = 10 mm. Percentage of Alizarin Red⁺ area in dish area. e Ratio of the expression of RUNX2, BGLAP, and SEMA3A as detected by RT-qPCR 1 week after the induction of osteogenesis. The results are shown as a ratio to the expression in MOCK-pretreated hSHED-EV-treated group (MOCK-EV = 1, respectively). f The expression of osteoblast markers, including RUNX2 and BGLA by Western blot analysis 1 week after the induction of osteogenesis. ACTB actin, beta. g The expression of SEMA3A in the conditioned medium by ELISA 1 week after the induction of osteogenesis. a–g MOCK-EV, MOCK-pretreated hSHED-EV-treated group; RNase-EV, RNase-pretreated hSHED-EV-treated group. n = 7 for all groups. Comparisons between two groups were analyzed by an independent two-tailed Student’s t test. Multiple group comparisons were performed by a one-way analysis of variance followed by Tukey’s post hoc test.*P < 0.05, ***P < 0.005. ns no significance. Graph bars represent mean ± SEM