Liver biopsies from humans without NASH (normal, n = 16) and with advanced NASH (NASH, n = 16) were examined for FOXO1 expression in hepatic macrophages. (A) Liver biopsies from normal subjects were stained with H&E, Oil Red O, and trichrome (original magnification, ×10). Scale bars: 100 μm. (B) Liver biopsies from patients with NASH were stained with H&E, Oil Red O, and trichrome (original magnification, ×10). Scale bars: 100 μm. (C) Hepatic TG content. (D) Hepatic cholesterol content. (E) Anti-FOXO1 and anti-F4/80 dual immunohistochemistry of liver biopsies from normal subjects (original magnification, ×20). Scale bars: 50 μm. (F) Anti-FOXO1 and anti-F4/80 dual immunohistochemistry of liver biopsies from patients with NASH (original magnification, ×20). Scale bars: 50 μm. (G) Hepatic mRNA levels of liver biopsies of normal and NASH subjects, as determined by real-time qRT-PCR assay. Data are expressed as mean ± SEM (n = 16). Statistical analysis in C was performed using a 2-tailed, unpaired t test, and in D and G using a 1-tailed, unpaired t test. *P < 0.05, **P < 0.01.