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. 2021 Jul 20;12:4404. doi: 10.1038/s41467-021-24601-y

Fig. 5. Autophagy induces fibroblast-to-myofibroblast transition, collagen release, and tissue fibrosis.

Fig. 5

a–c Cultured human dermal fibroblasts overexpressing BECLIN1. a Expression of BECLIN1 mRNA (n = 9 biological replicates per group) and protein (n = 5 biological replicates per group) in fibroblasts infected with adenoviral vectors encoding BECLIN1 or LacZ. b Effects of the adenoviral overexpression of BECLIN1 on the expression of αSMA and the formation of stress fibers (n = 8 biological per group) with representative images and quantification of fluorescence intensity. Fibroblasts stimulated with recombinant TGFβ served as positive control. Horizontal scale bar, 100 µm. c Changes in the mRNA levels of COL1A1 as analyzed by qRT-PCR (n = 4 biological replicates per group), on type I collagen protein as analyzed by western blot (n = 5 biological replicates per group) and on total collagen analyzed by hydroxyproline assays in fibroblasts upon adenoviral overexpression of BECLIN1 (n = 5 biological replicates per group). d, e Adenoviral overexpression of BECLIN1 in murine lungs. d Representative immunofluorescence staining of the markers of autophagy BECLIN1, ATG7, or p62 (all green) in combination with DAPI (blue) and vimentin (red) (n = 7 biological replicates per group). e Trichrome stainings, fibrotic area counts, myofibroblast counts, hydroxyproline content (n = 4 biological replicates for control group and n = 5 for AdBeclin1 group) and Ashcroft scores (n = 3 biological replicates for control group and n = 5 for AdBeclin1 group). Horizontal scale bars, 100 µm. All data are presented as median ± IQR. p-values were determined by two-sided Mann–Whitney test (a, d, e) or ANOVA one-way with Tukey’s multiple comparison post hoc test (b, c) and are indicated in the figure. See source data for more detailed information. rel.: relative, Ad: adenovirus, fluo.: fluorescence, int.: intensity, conc.: concentration.