Fig. 4. Hepatic PKD3 suppresses de novo lipogenesis in a SREBP-dependent manner.

A, B Basal and insulin-induced total de novo lipogenesis rate (measured under lipogenic conditions: serum deprived, 25 mM glucose, 0.5 mM sodium acetate, and 100 nM insulin) in primary hepatocytes isolated from mice of the indicated genotypes (arbitrary units – a.u.) (A) and quantification of TLC-separated free cholesterol (FC), triglycerides (TG), cholesteryl ester (CE), and other lipids (FFA, PE, and PL) (B) (n=3 biological replicates/group).

C In vivo de novo lipogenesis rate (a.u.) in livers from control and pKD3 ^liverΔ/Δ mice fed a HFD for 8 weeks, fasted overnight and re-fed for 4 hours before analysis (n=5 mice/group).

D Western blot analysis of mature SREBP1 in primary hepatocytes of the indicated genotypes under the same lipogenic conditions as B (n=3 mice/group; representative of three individual experiments).

E QPCR analysis of Srebp gene and target gene expression in primary hepatocytes of the indicated genotypes under the same lipogenic conditions as B (n=3 mice/group).

F QPCR analysis of Srebp gene and target genes expression in livers from control and pKD3 ^liverΔ/Δ mice fed for 20 weeks with HFD, subjected to fasting overnight and re-feeding for 4 hours (n=6 mice/group).

G Western blot analysis for indicated proteins in livers from the mice in F (n=3 mice/group).

H QPCR analysis of Srebp gene and target genes expression in primary hepatocytes of the indicated genotypes stimulated with insulin for 4 hours stimulation) that were either transfected with siNonTargeting (siNT) or the combination of slSrebp1 and slSrebp2 as indicated (n=3 biological replicates/condition).

Data information: In (A-C, E-F, and H), data are presented as mean ± SEM. *P>0.05, **P>0.01 (unpaired two-tailed Student’s t-test (C, E, F), or one way ANOVA with post hoc Tukey’s test (A, B, H)).