Research Paper Volume 16, Issue 5 pp 4095—4115

Figure 4. FoxO6 regulates hyperlipidemia through ApoC3 expression in FoxO6 virus-treated cells. (A) Expression of FoxO6 and ApoC3 by FoxO6. AC2F cells were grown to 80% confluence in 100-mm dishes using DMEM and then stimulated with 100 MOI of FoxO6 for 24 h and analyzed using western blotting using the appropriate antibody. Results are representative of three independent experiments. Bars in densitometry data represent the mean ± S.E., and significance was determined using one-factor ANOVA: ^*p < 0.05 vs. Normal. (B) Cellular TG contents were measured using a colorimetric assay. Results of one-factor ANOVA: ^***p < 0.001 vs. untreated cells. (C) FoxO6 binds to the ApoC3 promoter in liver cells. The cells were subjected to ChIP assay using rabbit pre-immune IgG and an anti-FoxO6 antibody. Immunoprecipitates were subjected to PCR using rat ApoC3 promoter-specific primers. (D) Cells incubated without or with FoxO6 (100 MOI) for 24 h were subjected to real-time PCR analyses to determine the mRNA levels of TG synthesis genes (MTP, ApoC3, ApoB, and ApoA1), lipogenesis genes (PPARγ, ACC, and FAS), and gluconeogenesis-related genes (PEPCK and G6Pase), using the β-actin gene as a control. Results of one-factor ANOVA: ^*p < 0.05, ^**p < 0.01, ^***p < 0.001 vs. untreated cells. (E) Predicted mechanism in aged liver tissues after HFD administration against lipid accumulation.