Figure 3.

Tn housing restricts WD + CCl₄ driven liver disease pathogenesis. (A) Schematic of eight-week-old WT mice maintained at Ts or acclimated to Tn for 2 weeks and treated with either WD + Corn Oil (control) or WD+CCL₄ 2µl (0.32 µg)/g of body weight i.p. once weekly for 12 weeks. During the course of the 12 weeks, (B) body weight was recorded. At the conclusion of the study, additional parameters of NAFLD severity were analyzed. (C) The liver tissue was preserved in formalin, stained with hematoxylin eosin (H&E), and analyzed by a clinical pathologist. Table depicting histological scoring analyses for fibrosis, lobular inflammation, hepatocellular ballooning, macrovesicular (MV) steatosis grade, steatosis percentage, portal inflammation and NAFLD activity score (NAS) severity. (D) Liver H&E staining. (Top and bottom) Predominant distribution of large- and small-droplet macrovesicular steatosis is in zone 1 (to 2), grade 2 (to 3) range, apparent at low power. Black bar = 336µm (green arrows: portal areas and cholangiolar proliferations; light blue arrows: central veins). Black bar = 115µm. (E) Liver H&E staining. (Top) Cholangiolar proliferations. Periportal multilayered and solid patches (double green arrows), focal multilayered pericellular (triple arrows). Black bar = 336µm. (Bottom) Extension to lobules (single arrows), focally zone 3 (near central vein), with associated large inflammatory focus in porta (blue arrow); nuclear atypia (orange arrow). Black bar = 38µm. (F) Serum ALT levels at the conclusion of the study. (G) Hepatic inflammation as defined by flow cytometric analyses of hepatic immune cell accrual. Donut charts depicting liver immune compositions (macrophages, neutrophils, CD4⁺ cells, CD8⁺ cells, NK cells, NKT cells, B cells, and unidentified cells as determined by flow cytometry from percent of CD45⁺ population. In bar graphs, data represent mean +/- SEM. (B, C, F, G) Data combined from 2 individual experiments (n=10-14/condition). (C, F) One way ANOVA. *P<.05, **P<.01.