proliferation and migration
(A) Representative transmission electron microscopy micrographs of PH rat plasma-extracted exosomes; (B) Analysis of exosomes size with Nanoparticle Tracking Analysis; (C) Detection of the exosomes marker CD9 with Western blotting (n=3); (D) The uptake of exosomes by PASMCs detected by confocal microscopy; (E)Immunofluorescence staining of α-SMA and PCNA in PASMCs; (F) The expression of α-SMA, SM22α and PCNA proteins in PASMCs analyzed by Western blotting (n=3); (G) EdU staining to evaluate PASMCs proliferation; (H) Flow cytometry to assess PASMCs proliferation; (I) PASMCs migration evaluated by scratch wound healing assay; (J) The representative immunoelectron microscopic images of exosomes. Immunostaining was performed using an anti-LOX-1 antibody (5 nm gold particles indicated by red arrows);(K) The expression of LOX-1 protein in plasma exosomes was analyzed by Western blotting (n=3); (L) Immunofluorescence staining of LOX-1 in PASMCs; (M) The expression of LOX-1 protein in PASMCs was analyzed by Western blotting (n=3).
Fig. 2. Plasma exosomes extracted from LOX-1-null PH rats blunted PASMCs phenotypic switching, proliferation and migration
(A) Immunofluorescence staining of α-SMA and PCNA in PASMCs;(B-C) The expression of α-SMA, SM22α and PCNA proteins in PASMCs was analyzed by Western blotting (n=3); (D) EdU staining to evaluate PASMCs proliferation; (E) Statistical summary of EdU staining (n=3); (F) Flow cytometry to evaluate PASMCs proliferation; (G) Statistical summary of flow cytometry (n=3);(H) PASMCs migration was evaluated by scratch wound healing assay; (I) Statistical summary of scratch wound healing assay (n=3).