Figure 5

LXR activation by TO901317 regulates anti-oxidant enzyme expression in Schwann cells. MSC80 cells were treated either with TO9 (10 µM) or vehicle (Ethanol) for 2 h, 4 h, and 8 h. Total proteins were extracted at the specified time points followed by Western Blotting. (a) Expression profile of Nrf2 protein quantified through western blots (cropped gels). All blots were normalized to alpha-tubulin expression. (b) Phosphorylation profile of Akt protein (Ratio P-Akt/Akt) quantified through western blots (cropped gels). All blots were normalized to alpha-tubulin expression. (c) mRNA expression profile of antioxidant response genes across 72 h. Transcript levels were normalized to GAPDH expression. Results represent the means ± SEM of at least three independent experiments. *P < 0.05, **P < 0.01 and ***P < 0.001 assessed by Tukey’s post hoc test after ANOVA. (d) Enzymatic activity of antioxidant response genes at 48 h and 72 h. Enzymatic activity is normalized to tissue weight. (e) MSC80 cells were transfected with either non-target siRNA (NT) or siRNA directed against LXRα and LXRß. 48 hours after transfection, cells were treated either with T09 (10 µM) or vehicle (Ethanol) for 24 h. mRNA levels of G6PDH, 6PGDH and IDH1 were assessed through RT-qPCR and normalized to GAPDH expression. Results represent the means ± SEM of at least three independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001 assessed by Student’s t test.