Panel A shows bar graphs with the y-axis representing Relative mRNA levels (fold changes) and the x-axis representing 0 d, 7 d, and 14 d for WT and Ifnlr1 minus/minus groups. Panel B shows histological images of kidney sections stained with Masson, PSR, alpha-SMA, and Fibronectin at 0 d, 7 d, and 14 d in WT and Ifnlr1 minus/minus mice. Panel C shows bar graphs with the y-axis representing Fibrotic area (percent) or Relative mean optical density and the x-axis representing 0 d, 7 d, and 14 d for WT and Ifnlr1 minus/minus groups. Panel D shows immunoblot images displaying alpha-SMA, Fibronectin, Vimentin, and GAPDH protein levels in kidney samples collected at 0 d, 7 d, and 14 d. Panel E shows bar graphs with the y-axis representing Relative mRNA levels (fold changes) and the x-axis representing 0 d, 7 d, and 14 d for WT and Ifnlr1 minus/minus groups. Panel F shows bar graphs with the y-axis representing Relative mRNA levels (fold changes) and the x-axis representing 0 d, 7 d, and 14 d comparing Mock and IFN-lambda2 groups. Panel G shows bar graphs with the y-axis representing Relative mRNA levels (fold changes) and the x-axis representing 0 d, 7 d, and 14 d in Renal TECs from Mock and IFN-lambda2 groups. Panel H shows histological images of kidney sections stained with Masson, PSR, alpha-SMA, and Fibronectin at 0 d, 7 d, and 14 d in Mock and IFN-lambda2-treated mice. Panel I shows bar graphs with the y-axis representing Fibrotic area (percent) or Relative mean optical density and the x-axis representing 0 d, 7 d, and 14 d comparing Mock and IFN-lambda2 groups. Panel J shows immunoblot images displaying alpha-SMA, Fibronectin, Vimentin, and GAPDH protein levels in kidneys from Mock and IFN-lambda2-treated mice at 0 d, 7 d, and 14 d. Panel K shows bar graphs with the y-axis representing Relative mRNA levels (fold changes) and the x-axis representing 0 d, 7 d, and 14 d comparing Mock and IFN-lambda2 groups.
IFN-λ triggers renal fibrosis in mice. (A–E) WT and Ifnlr1−/− mice underwent UUO surgery, and kidneys were harvested at days 0, 7, and 14 after surgery (n = 6). (A) RT-qPCR analysis for Isg15 and Mx1 mRNA levels in kidneys of WT and Ifnlr1−/− UUO mice at days 0, 7, and 14 after surgery. (B) Representative images showing Masson’s trichrome, PSR, α-SMA, and fibronectin staining in kidneys among groups (scale bars = 50 μm). (C) Quantitative analysis of B revealed the percentage of fibrotic area and α-SMA and fibronectin MOD in kidneys. (D) Western blot examination of α-SMA, fibronectin, and vimentin protein levels in kidneys from WT and Ifnlr1−/− UUO mice. GAPDH was employed as a loading control. (E) RT-qPCR analysis for renal Acta2, fibronectin, and vimentin mRNA levels in the indicated groups. (F–K) WT UUO mice were subcutaneously injected with 1 μg of IFN-λ2 on days −1, 1, 3, 5, 7, 9, 11, and 13. Kidneys and renal TECs were collected at days 0, 7, and 14 after UUO (n = 6). (F) RT-qPCR analysis for Isg15 and Mx1 mRNA levels in kidneys of IFN-λ2–treated UUO mice. (G) RT-qPCR analysis for Isg15 and Mx1 mRNA levels in renal TECs at indicated time points. (H) Representative images of Masson’s trichrome, PSR, α-SMA, and fibronectin staining in kidney sections at indicated time points (scale bars = 50 μm). (I) Quantitative analysis of H revealed differences in fibrotic area, α-SMA, and fibronectin MOD in the kidneys among groups. (J) Western blot assessment of α-SMA, fibronectin, and vimentin protein levels in UUO kidneys in the presence or absence of IFN-λ2 treatment. (K) RT-qPCR analysis of renal Acta2, fibronectin, and vimentin mRNA levels. Data in A, C, E–G, I, and K are pooled from two independent experiments. Data in B, D, H, and J are representative of two independent experiments. Data are shown as mean ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, by two-way ANOVA with Tukey’s multiple-comparison test. MOD, mean OD. Source data are available for this figure: SourceData F2.