Panel A shows flow cytometry plots for renal, skin, and peritoneal fibroblasts before and after purification. The axes are SSC-H and Vimentin, with percentages of Vimentin plus cells indicated. Panel B presents bar graphs of relative mRNA levels for Isg15, Mx1, Ifit1, and Oas1 in different fibroblast types treated with PBS or IFN-λ2. The axes are the gene names and fold changes. Panel C displays bar graphs of relative mRNA levels for the same genes in renal fibroblasts from WT and Ifnlr1 minus slash minus mice treated with PBS or IFN-λ2. The axes are the gene names and fold changes. Panel D shows a Western blot image assessing STAT1 phosphorylation and protein levels in renal fibroblasts from WT and Ifnlr1 minus slash minus mice treated with PBS or IFN-λ2. The lanes are labeled with treatment conditions, and GAPDH is used as a loading control.
IFN-λ acts on renal fibroblasts by activating the STAT1 signaling pathway. (A) Flow cytometric analysis of the proportions of renal fibroblasts, skin fibroblasts, and peritoneal fibroblasts before and after purification from different mouse tissues. Fibroblasts were gated on forward and side scatters, then single cell, live cells, and finally vimentin+ cells (n = 3). (B) Primary renal fibroblasts, skin fibroblasts, peritoneal fibroblasts, and MEFs were treated with 100 ng/ml of IFN-λ2 or PBS for 24 h. RT-qPCR was performed to determine Isg15, Mx1, Ifit1, and Oas1 mRNA levels (n = 4). (C and D) Primary renal fibroblasts isolated from WT and Ifnlr1−/− mice were treated with 100 ng/ml of IFN-λ2 or PBS for 24 h or 1 h for RT-qPCR and western blot analysis, respectively (n = 4). (C) RT-qPCR was performed to determine Isg15, Mx1, Ifit1, and Oas1 mRNA levels. (D) Western blot was used to assess STAT1 phosphorylation and protein levels. GAPDH was used as a loading control. Data are representative of two independent experiments. Data are presented as mean ± SEM. ****P < 0.0001, by unpaired two-tailed Student’s t test (A and B), or two-way ANOVA with Tukey’s multiple-comparison test (C and D). ns, no significant difference. Source data are available for this figure: SourceData FS2.