Fig. 3

solTNF neutralization decreases hepatic LCN2 in diet-induced liver inflammation. a HFHC diet impact on Srebp-1c mRNA expression (n = 6, diet effect P = 0.0263). b Increased hepatic triglyceride accumulation was observed in the HFHC groups (n = 8, diet effect P = 0.0102). c solTNF neutralization decreases elevated hepatic LCN2 associated with HFHC diet (n = 9–11, diet effect P = 0.0034, interaction P = 0.0034). d-g HFHC diet impact on Il-1 (n = 6, diet effect P < 0.0001), Il-6 (n = 6, diet effect P = 0.0036), Tlr2 (n = 5–6, diet effect P = 0.0221), and Ppar-α (n = 5–6, diet effect P = 0.0348) liver mRNA expression in the HFHC groups. h, i Immunoblot analysis demonstrates decreased IRS1 expression (n = 6, diet effect P = 0.0075) and phosphorylated of IRS1 (TYR608) (n = 6, diet effect P = 0.0372) after 14 weeks of HFHC diet treatment. j-l Scatter plots indicate significant Pearson correlation analysis (r value) between hepatic and colonic LCN2 levels (r² = 0.4650, P < 0.0002) and hepatic and plasma LCN2 (r² = 0.4168, P < 0.0001). Tissues were analyzed by qPCR using primers directed against murine Il-1, Il-6, Tlr2, and Ppar-alpha. RNA expression. For each animal, the Ct values were normalized to the Ct values for Gapdh and Ppia. The relative expression level of the target gene (fold change) was expressed as 2^−ΔΔCt, when compared with the mean DCt (threshold cycle) of the control group. Immunoblots are representative of two independent experiments. Band intensity was calculated using Image Studio Lite, and values were normalized to the intensity of β-actin. Blot images were cropped for comparison. Data were analyzed by two-way ANOVA followed by Tukey’s multiple comparisons in GraphPad Prism 6. Data in bar graphs are represented as the mean ± SEM. Lowercase letters indicate post hoc analysis. Values with different lowercase letters are significantly different from each other. Means with different lowercase letters are significantly different from each other, P < 0.05