Fig. 7: TNFAIP8-autophagy axis is associated with hepatic steatosis induced by EtOH.

a HCC cells were treated with 50 mM EtOH for 30 h, and lysates were immunoblotted with indicated antibodies. b HCC cells were transfected with EV or TNFAIP8-Myc plasmid for 18 h and treated with 50 mM EtOH for 24 h and lysates were immunoblotted indicated antibodies. c HCC cells were grown on a coverslip and transfected with EV or TNFAIP8-Myc plasmid for 18 h and treated with 50 mM EtOH and 100 µM OA for 24 h. Cells were stained with Oil Red O and TNFAIP8-Myc protein as described in materials and methods section and observed under an Olympus BX60 fluorescent microscope with the bright field (×40 objective) and photographed. Arrow indicates TNFAIP8 expression and cell steatosis. d Representative hematoxylin and eosin (H&E, original magnification ×400) stained liver sections of male C57BL/6J mice pair-fed chronic ethanol (EtOH) or control diets for 8 weeks. e C57BL/6J mice pair-fed chronic ethanol (EtOH) or control diets for 8 weeks, mice were euthanized, and 50 μg of liver lysates were immunoblotted with indicated antibodies (upper panel). * = non-specific band. Indicated protein levels from the upper panel were quantified using ImageJ software (https://imagej.nih.gov/ij/) and plotted (lower panel). ^$P < 0.05 relative to control mice. f TNFAIP8 expression is higher in hepatic steatosis human patients with a history of alcohol use: left and right panels: representative images and quantification of TNFAIP8 expression in liver tissue, including normal liver tissues, liver steatosis without alcohol use, and liver steatosis with a history of alcohol use. g Left panel: Representative images of TNFAIP8 expression in normal liver, steatohepatitis (NASH) with no history of alcohol use, and steatohepatitis with alcohol use. Right panel: the overall quantification of TNFAIP8 expression in normal liver tissue, liver steatosis without a history of alcohol use, and liver steatosis with a history of alcohol use. TNFAIP8 expression was analyzed and quantified from TMA. Boxplots show median and interquartile range with whiskers indicating the total range of TNFAIP8 expression. Data are expressed as the mean ± SEM  from 6 (e) mice or mean from 5–18 (f, g) human samples. *P < 0.05 relative to normal liver. ns: not significant.