Panel A shows vertical bar graphs comparing relative mRNA levels of Ifn-λ2, Ifn-λ3, Isg15, and Mx1 among different groups. The x-axis represents different treatment groups, and the y-axis represents relative mRNA levels. Panel B displays representative microscopy images of kidney sections stained with Masson's trichrome and PSR, along with bar graphs quantifying fibrotic areas. The x-axis shows different treatment groups, and the y-axis shows the percentage of fibrotic area. Panel C presents representative microscopy images of kidney sections stained for α-SMA, fibronectin, and p-SMAD2/3, along with bar graphs quantifying the relative mean optical density. The x-axis represents different treatment groups, and the y-axis shows the relative mean optical density. Panel D shows a Western blot analysis of alpha-SMA, fibronectin, vimentin, phosphorylated and total SMAD2/3 proteins in mouse kidneys. Panel E displays vertical bar graphs comparing relative mRNA levels of Acta2, fibronectin, and vimentin among different groups. The x-axis represents different treatment groups, and the y-axis represents relative mRNA levels. Panel F shows representative microscopy images of kidney sections stained with Masson's trichrome, PSR, and immunohistochemistry for alpha-SMA, fibronectin, and p-SMAD2/3, along with bar graphs quantifying fibrotic areas and relative mean optical density. The x-axis shows different treatment groups, and the y-axis shows the percentage of fibrotic area or relative mean optical density. Panel G presents a Western blot analysis of alpha-SMA, fibronectin, vimentin, phosphorylated and total SMAD2/3 proteins in mouse kidneys. Panel H displays vertical bar graphs comparing relative mRNA levels of Acta2, fibronectin, and vimentin among different groups. The x-axis represents different treatment groups, and the y-axis represents relative mRNA levels.
Neutralizing IFN-λ improves renal fibrosis in mice. (A–E) WT mice received intrarenal injections of an adenovirus vector expressing shRNA-targeting mouse IFN-λ2/3 (Sh-IFN-λ2/3) and control adenovirus-CMV-GFP (Sh-control) prior to sham or UUO surgery. Kidneys were harvested on day 7 after surgery. n = 6 per group. (A) RT-qPCR analysis for renal Ifn-λ2, Ifn-λ3, Isg15, and Mx1 mRNA levels among groups. (B) Representative images and quantitative analysis of fibrotic areas with Masson’s trichrome and PSR staining (scale bars = 50 μm). (C) Representative images and quantitative analysis of α-SMA, fibronectin, and p-SMAD2/3 MOD in mouse kidneys (scale bars = 50 μm). (D) Western blot examination of α-SMA, fibronectin, vimentin, phosphorylated and total SMAD2/3 proteins in mouse kidneys. (E) RT-qPCR analysis of Acta2, fibronectin, and vimentin mRNA levels in mouse kidneys. (F–H) WT UUO mice received intraperitoneal injections with anti-IFN–λ2/3 neutralizing antibody (anti–IFN-λ2/3) or IgG2b isotype control (anti-IgG2b) on days −1, 1, 3, and 5. The kidneys were collected at day 7 after UUO surgery (n = 6 per group). (F) Representative images and quantification analysis of fibrotic area, α-SMA, fibronectin, and p-SMAD2/3 using Masson’s trichrome, PSR, and IHC staining (scale bars = 50 μm). (G) Western blot analysis of α-SMA, fibronectin, vimentin, phosphorylated, and total SMAD2/3 proteins in mouse kidneys. (H) RT-qPCR analysis of Acta2, fibronectin, and vimentin mRNA levels in mouse kidneys. Data in A–H are representative of three independent experiments. Data are presented as mean ± SEM. **P < 0.01, ***P < 0.001, ****P < 0.0001, by two-way ANOVA with Tukey’s multiple-comparison test. ns, no significant difference. MOD, mean OD. Source data are available for this figure: SourceData F8.