Figure 3

EVs-HG suppressed the expression of TGF-β RII partly through transferring miR-17-5p. (A) TGF beta-related target genes for mmu-miR-17-5p were identified by the miRBase online database (https://www.mirbase.org). A higher score represents more statistical confidence in the prediction result. (B) Illustration of the potential base-pairing sites of miR-17-5p located in the 3’-UTR of TGF-β RII mRNA. (C) Analysis of TGF-β RII expression levels in MOVAS cells transfected with EVs-NG or EVs-HG and control mimic or miR-17-5p mimic. EVs-HG and miR-17-5p mimic significantly decreased TGF-β RII expression at transcript level respectively compared to EVs-NG or control mimic (N = 6). (D) The level of miR-17-5p was silenced in MOVAS cells by transfection of miR-17-5p inhibitor (N = 3). Inhibiting miR-17-5p in MOVAS cells significantly increased the TGF-β RII transcript levels compared to control inhibitor (N = 6). (E) EVs-HG and elevated expression of miR-17-5p reduced TGF-β RII expression at protein level respectively compared to EVs-NG or control mimic (N = 3). Original blots/gels are presented in Supplementary Figure S3−S4. (F) Schematic representation of the treatment of Fig. 3G is shown. (G) The gene expression of Runx2 was evaluated in the presence of 2 ng/ml TGF-β1. Inhibiting miR-17-5p in MOVAS cells cotransfected with EVs-HG significantly increased the Runx2 transcript levels (N = 6). GAPDH, β-actin, or U6 was used as the endogenous control. The p value was calculated by unpaired Student’s t test, and the data are presented as the mean ± SEM.