Fig. 3. Obese IFN-γ−/− mice develop accelerated NAFLD with fibrosis.

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Fig. 3. Obese IFN-γ−/− mice develop accelerated NAFLD with fibrosis. Obese IFN-γ−/− mice develop accelerated NAFLD with fibrosis. (A) Activation scores of the IFN-γ pathway were assessed by Ingenuity Pathway Analysis of significantly differentially expressed genes as determined by RNA-seq from whole liver tissue from WT, IL-4−/−, and IL-10/IL-4−/− mice on HFD for 15 weeks. Expression of genes annotated as being involved in positive regulation of IFN-γ is shown in the heat map. (B) WT (circles) and IFN-γ−/− (squares) mice were maintained for 15 weeks on a normal diet (filled symbols) or HFD (open symbols) and assessed for obesity by MRI body composition measurement (n = 3 to 5). NAFLD progression was also assessed by liver weight (n = 9 to 11), liver index, hepatic triglyceride measurement (n = 3 to 6) (B), steatosis by Oil Red O staining (C) (left, normal diet; right, HFD) (scale bars, 100 μm), picrosirius red staining of collagen (scale bars, 125 μm) (C) (bottom) (visualized under polarized light; inset scale bars, 125 μm), and serum ALT levels (n = 5) (D). (D) Expression of fibrotic markers col3a1 (n = 9 to 11), postn (n = 9 to 11), and acta2 (SD in shaded regions) (n = 3 to 5) was assessed by quantitative polymerase chain reaction (qPCR). (E) Expression of tgfb1 and TGF-β–regulated genes timp1 and mmp2 were assessed (n = 9 to 11). (F) Immunofluorescence staining for phospho-Smad3 (green), keratin 8/18 (gray), and 4′,6-diamidino-2-phenylindole (DAPI) (blue) (arrows indicate overlapping Smad3 and DAPI staining). All data points represent a single mouse, and representative or pooled data from two or more independent experiments are shown (two-tailed t tests, n = 2 to 10; *P < 0.05, **P < 0.01, ***P < 0.005, ****P < 0.0001). Kevin M. Hart et al., Sci Transl Med 2017;9:eaal3694 Published by AAAS