Figure 3
miR-13474 enriched in MSC-EVs and downregulated ADAM17 expression. (A) Relative percentage of miRNAs in total miRNA reads. (B) miR-13474 structure and sequencing results. (C) qRT-PCR to compare the expression of miR-13474 in extracellular vesicles of MSC and HFL (human embryonic lung fibroblasts) (n = 6). (D) qRT-PCR for miR-13474 in isolated MSCs and MSC-EVs (n = 3). (E) The predicted binding site of miR-13474 targeting the 3ʹ-UTR of ADAM17; Luciferase reporter assay showed ADAM17 as a target of miR-13474 (n = 6). (F) qRT-PCR of miR-13474 in EVs isolated from negative control (NC) mimics overexpressed MSC-EVs (miR-13474mimic NC-MSC-EVs) or miR-13474 mimics overexpressed MSC-EVs (miR-13474mimic-MSC-EVs) (n = 3). (G) Western blot detection of ADAM17 protein expression under miR-13474mimic-MSC-EVs treatment (n = 3). (H) qRT-PCR analysis of miR-13474 in EVs isolated from NC inhibitors transfected MSC (miR-13474inhibitor NC-MSC-EVs), or miR-13474 inhibitors transfected MSC (miR-13474inhibitor-MSC-EVs) (n = 3). (I) Western blot detection of ADAM17 under miR-13474inhibitor-MSC-EVs treatment (n = 3). All data are presented as means ± SEM. **P < 0.01 and ***P < 0.001. Original blots are presented in Supplementary Fig. S9.
