Fig. 5.

Phf2 and ChREBP decrease oxidative stress through Nrf2 activation. Mice, injected with either GFP or Phf2 overexpressing adenovirus, were studied 3 weeks later in fed state. a Representative Western blot analysis of Nrf2 protein content and Nrf2-regulated genes (n = 10 mice per group). b–d Nrf2 was inhibited in cultured hepatocytes overexpressing Phf2. b Western blot analysis of proteins involved in oxidative stress defenses (n = 3). GSH and NADPH contents (c) in addition to ROS levels and hepatocyte apoptosis (d) were determined after incubation with 480 μM palmitate for 24 h (n = 3). e UCSC genome browser image illustrating normalized tag counts for Phf2, ChREBP, H3K9me2, H3K4me3, and RNA polII at the Nrf2 promoter. (f, g) Phf2 was inhibited in cultured hepatocytes. ChIP for H3K9me2, ChREBP, and RNA polII, in addition to chromatin accessibility at the Nrf2 promoter and Nrf2 activity on the ARE-luc construct shown (n = 3). h–j ChREBP expression was inhibited in cultured hepatocytes overexpressing Phf2. h Representative western blot analysis showing the contribution of ChREBP to the regulation of Nrf2 and Nrf2-regulated gene expression. i Nrf2 activity on the ARE-luc construct shown. j Relative ROS levels and measurement of hepatocyte apoptosis were determined after incubation with 480 μM palmitate for 24 h (n = 3). All error bars represent mean ± SEM. Statistical analyses were made using Anova, followed by Bonferonni’s test. *P < 0.01 Phf2/USi compared to GFP/USi, **P < 0.01 Phf2/Nrf2i compared to Phf2/USi, ^#P < 0.01 GFP/USi palmitate compared to GFP/USi BSA, ^$P < 0.01 Phf2/USi palmitate compared to GFP/USi palmitate, ^£P < 0.01 Phf2/Nrf2i palmitate compared to Phf2/USi palmitate, a P < 0.01 USi 25 mM glucose compared to USi 5 mM glucose, b P < 0.01 Phf2i 25 mM glucose compared to USi 25 mM glucose, c P < 0.01 Phf2/USi compared to GFP/USi, d P < 0.01 Phf2/ChREBPi compared to Phf2/USi, e P < 0.01 GFP/USi palmitate compared to GFP/USi BSA, f P < 0.01 Phf2/USi palmitate compared to GFP/USi palmitate, and g P < 0.01 Phf2/ChREBPi palmitate compared to Phf2/USi palmitate