Fig. 4. High-fat and high-fructose diet induces liver inflammatory response.

(A) Surface staining of splenocytes, nodal immune cells, and hepatocytes for PDIA3 protein. (B) Normalized MFI of surface PDIA3 expressed on nodal immune cells, splenocytes, pancreatic cells, hepatocytes, liver immune infiltrates, cardiomyocytes, and adipocytes harvested from control and HFHF mice. A statistically significant difference in the PDIA3 HFHF/control ratio was observed. Values, from n = 4 biologically independent replicates, are reported as mean relative expression ± SD. Data were statistically analyzed using a two-tailed paired Student’s t test. Significance levels are reported as *P < 0.05. (C) Sequence of the linear autoantibody epitope (PDIA3 residues 104 to 128) shown in yellow mapped onto a surface representation of the PDIA3-tapasin complex crystal structure PDB.3F8U, with PDIA3 shown in magenta and tapasin shown in gray. (D to K) Livers were harvested from C57BL/6 mice on a regular diet or an HFHF diet for 3 months. Livers were fixed in formalin and stained to detect (D and E) steatosis (oil red staining), (F to I) hepatocyte injury and immune cell infiltration, and (J and K) proliferation. Steatosis, cellular infiltration and proliferation, lobular inflammation, and hepatocyte injury (ballooning) were all significantly up-regulated in HFHF mice as compared with controls. Values, from n = 6 biologically independent replicates, are reported as mean relative expression ± SD and were statistically analyzed using a two-tailed unpaired Student’s t test. (L and M) Increased ALT levels were seen in mice on a 3-month HFHF diet concomitantly with a decreased AST/ALT ratio (<1), which is indicative of NASH. Values from n = 13 biologically independent replicates are shown here, and data were statistically analyzed using a two-tailed unpaired Student’s t test.