Fig. 1: MV-induced pulmonary fibrosis was accompanied by fibrotic MV-EV release. | Experimental & Molecular Medicine

Fig. 1: MV-induced pulmonary fibrosis was accompanied by fibrotic MV-EV release.

From: ASK1-ER stress pathway-mediated fibrotic-EV release contributes to the interaction of alveolar epithelial cells and lung fibroblasts to promote mechanical ventilation-induced pulmonary fibrosis

Fig. 1

a Lung injury was assessed by hematoxylin and eosin staining. Collagen deposition was assessed with Masson’s trichrome staining. Original magnification ×200. Scale bars correspond to 100 μm (n = 6). b Fibrosis was also quantified by the determination of collagen-I α1 (COL1A1) and α smooth muscle actin (α-SMA) levels in lung tissues by western blotting (WB). Relative intensity of the protein bands of COL1A1 and α-SMA compared to that of Tubulin as determined by densitometry is displayed in bar graphs (n = 6). c Lung tissues were stained with fluorophore-labeled antibodies against COL1A1 and α-SMA (Alexa Fluor 594, red). 4’,6-Diamidino-2-phenylindole (DAPI) stain was used to detect nuclei (blue). Original magnification ×200. Scale bars correspond to 100 μm (n = 6). d EVs were quantified by protein BCA. e Confocal microscopy shows the fluorescence of uptake of PKH-67-labeled EVs (Alexa Fluor 488, green) by MRC-5 cells. Original magnification ×400. Scale bars correspond to 50 μm (n = 3). f Protein expression of COL1A1 and α-SMA in MRC-5 cells was determined by WB. Relative intensity of the protein bands of COL1A1 and α-SMA compared to that of Tubulin as determined by densitometry is displayed in bar graphs (n = 3). g GO enrichment analysis of DEGs between the EV(sham) and MV-EV groups. h Heatmap of upregulated and downregulated fibrosis-associated gene mRNA expression in the EV (sham) and MV-EV groups. The data are expressed as the means ± SEMs. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.001.

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