Figure 3

SCD1-mediated palmitoleate production enhances insulin-induced Wnt pathway activity in hepatocytes. Primary mouse hepatocytes were infected with Adv-TRE-Luc and Adv-RSV-β-gal and incubated in different conditions as indicated. Luciferase assays were performed and results are expressed as percent of the ratio firefly luciferase/β-galactosidase. (a) Adv-TRE-Luc-infected primary hepatocytes were incubated with palmitate, palmitoleate or control 0.6% BSA for 24 h as indicated. **p < 0.01, ***p < 0.001 vs BSA. (b) TRE-Luc-expressing primary hepatocytes were infected with either SCD1 or control GFP adenoviruses and incubated in the presence of palmitate (250 μM) or BSA for 24 h. Results are the mean ± SEM (n = 3). *p < 0.05, **p < 0.01 vs BSA; ^#p < 0.05 vs palmitate. (c) TRE-Luc-expressing primary hepatocytes were infected with GFP/USi, SCD1i or SCD1 adenoviruses and incubated in 25 mM glucose (G25) in the presence of the absence of insulin (ins) for 24 h. Results are the mean ± SEM (n = 3). ***p < 0.001 vs G25; ^#p < 0.05, ^###p < 0.001 vs G25 + ins. The Western blot shows SCD1 protein content in hepatocyte lysates from the different conditions. β-Actin antibody was used as a loading control. (d) RT-qPCR analysis of Scd1 and of Wnt target genes (GS and Lect2) expression in primary hepatocytes infected with the indicated adenoviruses and incubated in 25 mM glucose in the presence of the absence of insulin (ins) for 24 h. Results are the mean ± SEM (n = 3). **p < 0.01, ***p < 0.001 vs G25; ^#p < 0.05, ^##p < 0.01, ^###p < 0.001 vs G25 + ins.