Fig. 2.

Long-term HFD feeding induces GVB disruption and liver inflammation and steatosis. (A–H) Mice were fed with control (Ctrl) diet or HFD for 24 weeks before their intestines and livers were harvested. (A) Ileum and colon sections were stained for CD34 (green), PV1 (red) and DAPI (blue) expression, scale bar indicates 50 µm. (B) Quantification of PV1 MFI was performed on CD34⁺ area. (C) LPS levels were measured in the serum of Ctrl or HFD-fed mice. (D) Liver sections were analyzed by H&E, ORO or Sirius Red staining, as indicated. Scale bar indicates 100 µm. (E) ALT serum concentration in Ctrl or HFD-fed mice. (F) Glucose tolerance test and insulin tolerance test were performed after 6 h of fasting and intraperitoneal injection of glucose (2 g/kg mouse) and insulin (0.2 IU/kg mouse) respectively. (G) Analysis of gene expression (normalized to PPIA gene) by qPCR in the liver of Ctrl or HFD-fed mice. (H) FACS staining of immune cells in the liver of Ctrl or HFD-fed mice represented as absolute numbers. *p <0.05; **p <0.005; ***p <0.0005; unpaired 2-tailed t test or 2-way ANOVA in panel F. ALT, alanine aminotransferase; GVB, gut-vascular barrier; HFD, high-fat diet; LPS, lipopolysaccharide; MFI, mean fluorescence intensity; ORO, Oil Red O.