Fig. 6: Idh3 overexpression ameliorates axonal ATP deficits in EAE lesions.
a, Experimental design to measure [ATP/ADP]axon in acute EAE in Thy1-PercevalHR mice that virally overexpressed Idh3a or a control protein (Cre recombinase) together with tdTomato in a subset of axons. b, Maximum intensity projections of in vivo multi-photon image stacks of spinal cord axons in Idh3a-overexpressing Thy1-PercevalHR mice. Grayscale LUT of tdTomato (left). Ratiometric [ATP/ADP]axon LUT (λex ratio 950 nm/840 nm) (right). Details below show image pairs of tdTomato-negative (tdTom−, left) and tdTomato-positive (tdTom+, right) normal-appearing, swollen and fragmented axons (tdTom− has dashed outlines) in acute EAE. c, Comparison of [ATP/ADP]axon in tdTom+ and tdTom− axons (plotted as λex ratio 950 nm/840 nm, normalized to control axon mean indicated as the black line; values above 1.5 are lined up on the ‘≥1.5’ dashed line). [ATP/ADP]axon of single tdTom− (gray) and tdTom+ (orange) axons in Idh3a-overexpressing EAE mice (left). Lesion-specific paired analysis of mean [ATP/ADP]axon in tdTom− (gray) and tdTom+ (orange) axon populations of the three morphological stages (right). d, Same analysis as c, but in a mouse cohort overexpressing a control protein (Cre recombinase). Mean ± s.e.m. Comparison of 176 tdTom− axons versus 153 tdTom+ axons in 12 lesions from four mice in c; 214 tdTom− axons versus 159 tdTom+ axons in 11 lesions from three mice in d using a two-tailed, unpaired Student’s t-test (c,d, left graphs) and a paired t-test (c,d, right graphs). Scale bar, 25 μm (b). **P < 0.01; ***P < 0.005; ****P < 0.001. See source data for individual data points and further statistical parameters. Illustration created with BioRender.
