External factors
| External factors: |
Exosome |
| Category: |
Chemical compounds |
| Experiment: |
TEM//SA-β-gal activity assay//Western blot//Flow cytometry |
| Description: |
Western blot analysis confirmed that the isolated particles expressed exosome-specific markers: CD63, CD9, and Alixs. The particle size of ExoUMSCs was around 60–150nm as shown by dynamic light scattering (DLS). After OMSCs were incubated with Dil-labeled ExoUMSCs for 48h, fluorescence was detected inside the cells, indicating that ExoUMSCs?were efficiently internalized into the target cells. A number of β-gal-positive cells were significantly reduced in OMSCs after treated with ExoUMSCs. Expression of aging-related factors p53, p21, and p16 was also markedly reduced, and the level of Sirt1 was increased in the ExoUMSCs-treated OMSCs. The growth rate of OMSCs was significantly increased, and more OMSCs entered into S phase of cell cycle after treatment with ExoUMSCs. The numbers of EdU-positive OMSCs were significantly higher in the ExoUMSCs-treated OMSCs than the untreated cells. |
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Regulatory relationship
| R-EF-Target gene:
|
Upregulation |
| Official symbol(s): |
MIRNA-136 |
| Target gene experiment: |
qRT-PCR//Western blot |
| Target gene description: |
Among these miRNAs, miR-136 was the most abundant in UMSCs, as compared with OMSCs, and was increased more than 20-fold in OMSCs after cultured with ExoUMSCs, whereas no significant difference was observed for other miRNAs such as miR-106a, miR-155, and miR-29C among these groups. Moreover, the level of miRNA-136 in ExoUMSCs was also higher than that in ExoOMSCs. Next, we examined the role of miR-136 in regulating OMSC senescence and biological function. The mRNA and protein levels of senescent-associated markers p53, p21 and p16, as well as the β-gal+?cells, were markedly reduced in OMSCs after transfected with miR-136 mimics. |
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Aging network
Annotation:
The green line represents Upregulation.
The purple line represents Downregulation.
The orange line represents Activation.
The yellow line represents Inhibition.
The gray line represents Unclear.
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