Figure 5. The Sema7a^R145W mutation enhances hepatic FA and TG synthesis and FA uptake by enhancing PKC-α signaling–stimulated expression of transcription factors Srebp1 and Chrebp and nuclear receptor Lxr.

(A) Western blot analysis of the relative levels of Srebp1, Chrebp, Lxr, phosphorylated PKC-α, and PKC-α protein expression in 10-week-old male WT mice (n = 4), Sema7a^R145W heterozygous mice (n = 5), and Sema7a^R145W homozygous mice (n = 5). Western blot analysis of the relative levels of nuclear Srebp1, Chrebp, and Lxr proteins in primary hepatocytes from WT and Sema7a^R145W homozygous mice (B) and in human hepatoma HepG2 cells (C) after transfection with the plasmid for the expression of SEMA7A_WT and SEMA7A_R148W proteins. (D) Representative Western blot of the relative levels of nuclear Srebp1, Chrebp, and Lxr proteins in nuclear extracts and (E) Fasn, Acc1, and Cd36 proteins in whole-cell lysates of primary mouse hepatocytes after transfection with empty vector (CTR) or the plasmid for the expression of PKCα_WT or PKCα_dominant negative (DN) mutant, respectively. All primary mouse hepatocytes were isolated from 12-week-old male WT and Sema7a^R145W homozygous mice. Data are representative images or expressed as the mean ± SD of each group from 3 separate experiments. The data were analyzed by 1-way ANOVA with Tukey’s post hoc tests or by Kruskal-Wallis test with Dunn’s post hoc test analysis. *P < 0.05 versus the WT mice, ^#P < 0.05 versus the Sema7a^R145W heterozygous mice, ^$P < 0.05 versus the primary HO mouse hepatocytes transfected with CTR; n = 3.