Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Hepatocyte Toll-Like Receptor 5 Promotes Bacterial Clearance and Protects Mice Against High-Fat Diet–Induced Liver Disease

doi: 10.1016/j.jcmgh.2016.04.007

Figure Lengend Snippet: Liver TLR5 protects against high-fat diet–induced fibrosis. WT, TLR5KO, TLR5 fl/fl , TLR5 ΔHep , and TLR5-NLRC4 double knock-out (DKO) mice were fed with a regular chow diet or a HFD, comprising 60% fat, for 8 weeks. ( A and B ) Liver fibrosis estimation using Sirius red staining, with ( A ) representative images and ( B ) quantification shown. ( C ) Liver fibrosis estimation using Masson's trichrome staining, with representative images shown. ( D ) Hydroxyproline quantification. ( E and F ) Liver fibrosis estimation using collagen immunostaining and confocal microscopy analysis, with ( E ) representative images (staining of collagen in red and DNA in blue), and ( F ) quantification (percentage of vein presenting collagen accumulation, determined through the examination of 10 veins per slide) shown. ( G ) Liver and ( H ) adipose tissue mRNA were isolated and quantitated for expression of the CXCL1 gene, involved in inflammation. Results are expressed as relative values compared with WT mice fed with regular chow, defined as 1. ( I ) Analysis of CXCL1 protein expression level by enzyme-linked immunosorbent assay in the serum. ( J ) Analysis of IL1β protein expression level by enzyme-linked immunosorbent assay in the serum. Scale bar : 50 μm. Data are the means ± SEM. N = 5. Significance was determined by the Student t test. * P < .05.

Article Snippet: Tissues were sectioned at 5-μm thickness and subjected to collagen immunostaining (primary antibody: anti-mouse collagen type I, AB765P; Millipore; secondary antibody: anti-rabbit IgG cy5) with DNA staining using Hoechst.

Techniques: Knock-Out, Staining, Immunostaining, Confocal Microscopy, Isolation, Expressing, Enzyme-linked Immunosorbent Assay