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Fig. 6 | Stem Cell Research & Therapy

Fig. 6

From: Exosomes derived from human umbilical cord MSCs rejuvenate aged MSCs and enhance their functions for myocardial repair

Fig. 6

MiR-136 downregulates Apaf1 expression through directly binding with its noncoding region. a Real-time RT-PCR analysis of Apaf1 level in OMSCs and UMSCs, GAPDH used as control. b. Western blot analysis of senescence-related proteins p53, p21, and p16 in OMSCs treated with agonist (MDK83190) and inhibitor (ZYZ-488) of apaf1. GAPDH used as control. c Real-time RT-PCR analysis of Apaf1 level in OMSCs which were treated with ExoUMSCs or transfected with miR-136 mimic or miR-NC for 48 h (left panel), or Apaf1 mRNA in UMSCs after the cells were transfected with miR-136 inhibitor (anti-miR-136) or anti-miR-NC for 48 h (right). d Apaf1 was predicted as the target mRNA of miR-136 by TargetScan (right). Schematic diagram showed the potential two binding sites for miR-136 on the 3′UTR (138–145) and (1091–1098) of Apaf1, and their designed mutants. Luciferase reporter vectors with different Apaf1 3′UTR (WT, double mutations at both sites (D-MUT), single mutation at either site (S-MUT1 and S-MUT2) were constructed (left). e 293 T cells were transfected with the specified luciferase reporter vector containing the WT, mutant plasmids S-MUT1, S-MUT2, and D-MUT, along with miR-136 mimic or miR-NC. After 24 h, cell lysates were prepared and subjected to determination of relative luciferase activity by normalization of firefly luciferase activity with Renilla luciferase activity. f Western blot analysis of Apaf1 and its downstream target caspase 9 protein in OMSCs which were transfected with miR-136 mimic or miR-NC. GAPDH used as control. g Schematic cartoon illustrating that exosomes derived from human umbilical cord MSCs rejuvenate aged MSCs and enhance their functions for myocardial repair. *P < 0.05 vs OMSCs/UMSCs+miR-NC; *P < 0.05 vs DMEM

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