Endothelial MicroRNA-483-3p Is Hypertension-Protective

Figure 4

EC-SMC cross talk via exosome-delivered miR-483-3p. (a) A representative electron microscopy image of HUVEC-derived exosomes. Scale bar, 100 nm. (b) A nanoparticle tracking analysis of the size distribution of exosomes. (c) Immunofluorescence images of HASMCs stained with α-SMA (red) and DAPI (blue). HASMCs were treated with HUVEC-derived exosomes (PKH67-label, green) for 0 or 24 hours. Laser scanning confocal images are shown. Scale bar, 20 μm. (d) The qPCR analysis of miR-483-3p levels in HUVECs transfected with pre-miR-Ctrl or pre-miR-483-3p for 48 hours. (e) Exosomes isolated from HUVECs were identified by CD81 and CD63 immunostaining. (f) miR-483-3p levels in exosomes isolated from HUVECs transfected with pre-miR-Ctrl or pre-miR-483-3p. (g–i) HASMCs were incubated with HUVEC-derived exosomes for 24 hours. The qPCR analysis of (g) miR-483-3p and ACE1, TGF-β, and (h) CTGF in HASMCs. (i) Western blotting analysis of these proteins in HASMCs. Data are shown as . Student’s two-tailed test was used for statistical analysis of data. ; .

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