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. 2023 Feb 15;15(1):mjad009. doi: 10.1093/jmcb/mjad009

Figure 4.

Figure 4

Blocking autophagy aggravates TGF-β1-induced fibrosis in primary ConFB. (A) Knockdown of ATG3 or ATG7 using lentiviral-shRNA resulted in autophagy blockage and accumulation of SQSTM1, confirmed by immunoblots (left). Quantified protein levels were displayed as bar graphs (right). Data are presented as mean ± SD from three independent experiments (*P < 0.05, ****P < 0.0001 vs. normal control group; one-way ANOVA followed by Bonferroni post hoc test). (B–D) Primary ConFB before and after ATG3 or ATG7 knockdown were incubated in the presence or absence of 1 ng/ml TGF-β1 for 48 h. (B) Immunoblots showing protein levels of α-SMA and FN (left). Quantified protein levels were displayed as bar graphs (right). (C) Viability determined by the CCK8 assay. (D) Invasive ability determined by the transwell assay (left). Scale bar, 50 μm. The average numbers of invading cells per visual field were displayed in a bar graph (right). Data are presented as mean ± SD from three independent experiments (*P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001 vs. normal control group (B and C) or scramble control group (D); one-way ANOVA followed by Bonferroni post hoc test). (E) Immunoblots showing protein levels of LC3, α-SMA, FN, BECN1, ULK1, and WIPI1 in primary ConFB before and after knockdown of BECN1, ULK1, or WIPI1 incubated in the presence or absence of 1 ng/ml TGF-β1 for 48 h (left). Quantified protein levels were displayed as bar graphs (right). Data are presented as mean ± SD from three independent experiments (****P < 0.0001 vs. normal control group; one-way ANOVA followed by Bonferroni post hoc test). SC, scramble control; KD, knockdown.