Research Paper Volume 16, Issue 5 pp 4095—4115

Figure 3. High-glucose induced hyperlipidemia through FoxO6-mediated ApoC3 expression. (A) Western blot was used to detect p-FoxO6, FoxO6 in nuclear extracts, and ApoC3 in cytoplasmic extracts after treatment of AC2F cells with glucose (25 mM) for 6 h. TFIIB was the loading control of the nuclear fractions, whereas β-actin was the loading control of the cytosolic fractions. Results are representative of three independent experiments. Bars in the densitometry data represent the mean ± S.E., and significance was determined using one-factor ANOVA: ^**p < 0.01, ^***p < 0.001 vs. Normal. (B) Immunohistochemical staining for FoxO6 in cells with high-glucose treatment. Scale bar: 100 μm. (C) Cellular triglyceride contents after glucose treatment (25 mM) for 24 h was measured using a colorimetric assay. Results of one-factor ANOVA: ^***p < 0.001 vs. non-treated cells. Three independent experiments were performed and similar results were obtained. (D) Real-time PCR analyses were conducted for measuring the mRNA levels of the lipoprotein metabolism-related genes (MTP, ApoC3, ApoB, and ApoA1), lipogenesis genes (PPARγ, FAS, and ACC) and gluconeogenesis-related genes (PEPCK and G6Pase). Three independent experiments were performed and similar results were obtained. Results of one-factor ANOVA: ^*p < 0.05, ^**p < 0.01, ^***p < 0.001 vs. non-treated cells.