Fig. 4: Tumor and diet-induced hepatic insulin resistance, impaired lipid metabolism, and liver-WAT crosstalk.

To evaluate the insulin signaling pathway, a proportion of the animals received an insulin shot before euthanasia as described in the Materials and Methods section. A Representatives immunoblot analysis of insulin receptor β (IRβ) expression, phosphorylated AKT (T308 and S473), mTOR (S2448), and Ribosomal protein S6 kinase beta-1 (p70S6K) (T421/S424) expression. The total proteins or β-actin were used as loading control. B Representative immunoblotting of phosphorylated PKC (Thr514) and total PKC in liver lysates (GAPDH was used as loading control). C Representative immunoblotting of phosphorylated ATP citrate lyase (ACLy) (S455), acetyl-CoA carboxylase (ACC) (S79), and total ACLy, ACC, and stearoyl-CoA desaturase 1 (SCD1) in liver lysates (HSP90 was used as loading control). The results represent three independent experiments (n = 3). Western blot densitometries are in S4. D Scd1. E Fibroblast growth factor 21 (Fgf21). F Spliced form of X-box binding protein 1 (Xbp1s). G ApoB mRNA expression. H Cd36. I Peroxisome proliferator-activated receptor gamma coactivator 1-alpha (Pgc1a). J Cpt1a and K Acyl-CoA dehydrogenase long chain (Acadl) mRNA expression in liver from CR, Chow, or HFHS fed animals from all groups. Plotted values are mean ± S.E.M. ^*P < 0.05; ^**P < 0.01; ^***P < 0.001; ^****P < 0.0001. For all panels, a two-way ANOVA was performed, followed by Tukey’s post-test. qPCR reactions for each sample were performed in duplicate. All liver samples were analyzed.