Fig. 5
Effects of miR-136 on OMSCs. a Real-time RT-PCR analysis of miR-136 expression in UMSCs and OMSCs with or without treatment with ExoUMSCs. b Real-time PT-PCR analysis of miR-136 levels in exosomes derived from OMSCs and UMSCs. The expression level of miR-136 was normalized to U6. c Real-time RT-PCR analysis of p53, p21, and p16 expressions in OMSCs which were transfected with miR-136 mimic or scrambled miRNA (miR-NC), or treated with ExoUMSCs for 48 h. d Western blot analysis of senescence-related proteins p53, p21, and p16 in OMSCs transfected with miR-136 mimic or miR-NC. GAPDH was used as a loading control. e Representative images of β-gal staining in OMSCs transfected with miR-136 mimic or miR-NC. β-gal-positive cells out of total cells were quantified in the bar graph. f Representative images of immunofluorescent-stained OMSCs which were transfected with miR-136 mimic or miR-NC, or treated with ExoUMSCs for 48 h, and then stained with Ab against γ-H2AX (red) and DAPI (blue). Scale bar, 100 μm. Quantification of γ-H2AX foci in the specified cells. g Quantification of Edu-positive cells in OMSCs which were treated with ExoUMSCs, or transfected with miR-136 mimic or miR-NC. h Quantification of OMSCs at S phase of cell cycle as a percentage of total cells. OMSCs were cultured under the specified conditions and analyzed for their DNA contents using flow cytometry. i Quantification of apoptotic OMSCs which were treated with ExoUMSCs or transfected with miR-136 mimic or miR-NC, and then cultured under hypoxia and serum deprivation conditions. Apoptosis of OMSCs was detected by Annexin V/PI staining. j Cell viability of OMSCs with indicated treatments were assessed by CCK-8 assay under serum deprivation under hypoxia condition. *P < 0.05 vs OMSCs/DMEM; **P < 0.05 vs OMSCs; #P < 0.05 vs OMSCs + miR-NC/miR-136