Figure 6.

Insulin resistance does not impair overload-stimulated activation of the pentose phosphate pathway. After 12 weeks of high-fat diet (HFD) feeding, mice underwent unilateral synergist ablation surgery to induce overload (OVL) of the plantaris muscle for 5 days. (A) Muscles were excised and the protein content of glucose-6-phosphate dehydrogenase (G6PD) determined by immunoblot (IB) analysis. The G6PD antibody was validated using mouse tibialis anterior muscle samples overexpressing G6PDx (OE) or empty vector-transfected controls (-). (B–D) Ultra performance liquid chromatography/ multiple reaction monitoring-mass spectrometry (UPLC/MRM-MS) was used to measure 6-phosphogluconate, NADP and NADPH. (E) The ratio of NADP:NADPH was calculated. Data are mean ± standard deviation. Statistical analysis was performed using a 2-way ANOVA with Tukey’s post-hoc analysis. N = 7–9 muscles/group. p < 0.05 # vs. Sham (Sh); * vs. low-fat diet (LFD).