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. 2018 Nov 12;2018:6957497. doi: 10.1155/2018/6957497

Figure 2.

Figure 2

Stronger proinflammatory immune response and fibrosis development in livers of LysCre/c-Metmut animals. (a) Intrahepatic CD4+ and CD8+ T cells were analyzed by flow cytometry after chow, 4 weeks of MCD, or 24 weeks of HFD feeding of c-Metfl/fl and LysCre/c-Metmut mice. CD4+ and CD8+ T cells were gated by FSC/SSC (duplets were excluded), live/CD45+, CD4+ , or CD8+. A statistical analysis of the ratio of CD4+/CD8+ T cells of recorded cells was performed (n = 5) (∗ p < 0.05, ∗∗∗ p < 0.001). (b) mRNA expression levels of TNF-α, IL-6, TGF-β, and Collagen1α. Whole liver homogenates of c-Metfl/fl or LysCre/c-Metmut mice were analyzed via real-time PCR. The quantification is expressed as fold induction over the mean values obtained for chow fed c-Metfl/fl livers. At least 6 animals per group were included (∗ p < 0.05, ∗∗∗ p < 0.001). (c) Statistical analysis of the percentage of TUNEL+ and DHE+ cells referred to the number of total cells on stained liver sections of c-Metfl/fl and LysCre/c-Metmut mice treated either with chow or steatohepatitis-induced diets. 10 view fields/liver of at least n = 4 animals per genotype and time point were included (scale bars: 100 μm; magnification: 200x) (∗ p < 0.05). (d) Quantitative analysis of α-SMA-stained liver cryosections of c-Metfl/fl and LysCre/c-Metmut mice after chow, MCD (4 weeks), or HFD (24 weeks) feeding. Scale bars: 100 μm; magnification: 200x. Quantitative analysis of α-SMA staining was calculated by evaluating the α-SMA+ area per field area of 10 view fields/liver by ImageJ© (∗ p < 0.05). n = 4 animals per group and genotype were included. (e) For quantitative analysis of Sirius Red staining, the Sirius Red-positive area per view field of 10 view fields/liver of chow, MCD (4 weeks), or HFD (24 weeks) fed c-Metfl/fl or LysCre/c-Metmut animals was analyzed and recorded under polarized light by ImageJ© (∗ p < 0.05, ∗∗ p < 0.01). Included were at least n = 4 animals per genotype and time point. (f) Displayed are hydroxyproline levels of chow, MCD (4 weeks), or HFD (24 weeks) fed c-Metfl/fl and LysCre/c-Metmut mice (∗ p < 0.05) (n = 4).