Research Paper Volume 16, Issue 5 pp 4095—4115

Figure 2. Aging-related increase in hyperlipidemia through FoxO6-induced apolipoprotein expression. (A) Western blotting was performed to investigate the protein expression levels of p-FoxO6 and FoxO6 in the liver of aged rats. TFIIB was the loading control of the nuclear 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 < 0.05, ^##p < 0.01, ^###p < 0.001 vs. Young; ^***p < 0.001 vs. HFD-Young; ^$$p < 0.01 vs. Old. (B) Immunohistochemical staining for FoxO6 in the liver of aged rats. Scale bar: 100 μm. (C) Western blotting was performed to investigate the protein levels of ApoC3 and ApoB in the liver of aged rats. β-actin was the loading control of the cytosolic fraction. Bars in the densitometry data represent the mean ± S.E., and significance was determined using one-factor ANOVA: ^##p < 0.01, ^###p < 0.001 vs. Young; ^***p < 0.001 vs. HFD-Young; ^$$$p < 0.001 vs. Old. (D) Real-time PCR analyses were performed to measure the mRNA levels of FoxO6, MTP, ApoC3, ApoB, and ApoA1. Results of one-factor ANOVA: ^*p < 0.05, ^**p < 0.01, ^***p < 0.001 vs. Young; ^#p < 0.05, ^##p < 0.01 vs. Young-HFD. (E) Hepatic TG in HFD-fed aged rats. Results of one-factor ANOVA ^*p < 0.05, ^**p < 0.01 vs. Young; ^#p < 0.05 vs. Old. (F) Real-time PCR analyses were performed for measuring the mRNA levels of PPARγ, ACC, and FAS. Results of one-factor ANOVA: ^*p < 0.05, ^**p < 0.01 vs. Young; ^#p < 0.05, ^##p < 0.01 vs. Young-HFD. (G) G6Pase and PEPCK mRNA levels (gluconeogenesis-related genes) in the livers of HFD-fed aged rats. Results of one-factor ANOVA: ^*p < 0.05 vs. Young; ^#p < 0.05, ^##p < 0.01 vs. Young-HFD. (H) Plasma levels of ApoC3 were determined. Results of one-factor ANOVA: ^**p < 0.01 vs. Young; ^#p < 0.05 vs. Young-HFD.