Fig. 5

Immunofluorescence staining for GLUT4 protein in the skeletal muscle of WT and KO mice. Ten micrometer, fresh-frozen sections from KO and WT mice fed a high-fat or chow diet for 8 weeks were incubated overnight with rabbit anti-mouse GLUT4 antibody followed by incubation with an Alexa Fluor®568-conjugated goat anti-rabbit IgG secondary antibody followed by mounting in fluorescence mounting medium containing DAPI. Representative images from 3 WT and 4 KO mice per group are shown at 40X magnification. Primary antibody was omitted from the unstained control. Mean fluorescence intensity of 20 myofibers from 2 sections per animal was quantified using Image J. **P < 0.01; ***P <0.001 by 2-way ANOVA followed by Bonferroni post-hoc tests. ^† P < 0.05 for the interaction of diet and genotype by 2-way ANOVA