c-Met Signaling Protects from Nonalcoholic Steatohepatitis- (NASH-) Induced Fibrosis in Different Liver Cell Types

Figure 5

Loss of c-Met on MxCre⁺ cells strengthens the proinflammatory immune response after chronic liver injury. (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-Met^fl/fl and MxCre/c-Met^mut 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 () (). (b) mRNA expression levels of IL-6, TGF-β, Collagen1α, and α-SMA. Whole liver homogenates of c-Met^fl/fl or MxCre/c-Met^mut mice were analyzed via real-time PCR. The quantification is expressed as fold induction over the mean values obtained for chow fed c-Met^fl/fl livers. At least 5 animals per group were included (). (c) Statistical analysis of the percentage of TUNEL⁺ and DHE⁺ cells referred to the total number of cells on stained liver sections of c-Met^fl/fl and MxCre/c-Met^mut mice treated either with chow or steatohepatitis-induced diets. 10 view fields/liver of at least animals per genotype and time point were included (scale bars: 100 μm; magnification: 200x) (, ). (d) Quantitative analysis of α-SMA-stained liver cryosections of c-Met^fl/fl and MxCre/c-Met^mut 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© (). 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-Met^fl/fl or MxCre/c-Met^mut animals was analyzed and recorded under polarized light by ImageJ© (, ). Included were at least animals per genotype and time point. (f) Hydroxyproline levels of chow, MCD (4 weeks), or HFD (24 weeks) fed c-Met^fl/fl and MxCre/c-Met^mut mice are displayed ().

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