Fig. 5

Extracellular vesicles from HBE cells transfer miR-422a to MRC-5 cells, regulating myofibroblast differentiation phenotype. Note: ExoQuick-TC was used to isolate extracellular vesicles from normal or CSE-treated HBE cells. (A) Transmission electron microscopy images of extracellular vesicles (bar = 100 nm); (B) Particle number and size of HBE-exo or CHBE-exo were measured using the ZetaView® nanoparticle tracking analyzer based on dynamic light scattering; (C) Expression of CD9, Alix, and GRP94 in extracellular vesicles was examined by Western blot; (D) The levels of miR-422a in extracellular vesicles from normal or CSE-treated HBE cells were detected by qRT-PCR. (E) Microscopic images of MRC-5 cells incubated with PKH67-labeled (green) extracellular vesicles, with cell nuclei stained with DAPI (blue); (F) Co-culture of MRC-5 cells with extracellular vesicles (50 µg/mL) from normal or CSE-treated HBE cells, with expression levels of miR-422a, SPP1, and IL17A in MRC-5 cells detected by qRT-PCR; (G) Relative protein levels of α-SMA and type I collagen in MRC-5 cells were measured by Western blot. HBE-EVs, extracellular vesicles derived from normal HBE cells; CHBE-EVs, extracellular vesicles derived from CSE-treated HBE cells. Experiments were repeated at least three times. * indicates P < 0.05