Extended Data Fig. 7: Mdh2 overexpression ameliorates axonal ATP deficits in EAE lesions.
(a) Experimental design for [ATP/ADP]axon in acute EAE in Thy1-PercevalHR mice that virally overexpressed Mdh2 together with tdTomato in a subset of axons. (b) Maximum intensity projections of in vivo multi-photon image stacks of spinal cord axons in Mdh2-overexpressing Thy1-PercevalHR mice. Left: Grayscale LUT of tdTomato. Right: Ratiometric [ATP/ADP]axon LUT (λex ratio 950 nm/840 nm). Details show image pairs of tdTomato-negative (left) and -positive (right) normal-appearing, swollen, and fragmented axons in acute EAE. (c) Comparison of [ATP/ADP]axon in tdTomato-positive (tdTom+) and -negative (tdTom−) axons (plotted as λex ratio 950 nm/840 nm, normalized to control axon mean indicated as the black line; values above 1.5 lined up on the “≥1.5” dashed line). Left: [ATP/ADP]axon of single tdTom− (gray) and tdTom+ (orange) axons in Mdh2-overexpressing EAE mice, mean ± s.e.m. of tdTom+ stage 2 axons are above 1.5. Right: Lesion-specific paired analysis of mean [ATP/ADP]axon in tdTom− (gray) and tdTom+ (orange) axon populations of the three morphological stages. Mean ± s.e.m. Comparison of 110 tdTom−axons versus 112 tdTom+ axons in 14 lesions from four mice in using two-tailed, unpaired Student’s t-test or Mann-Whitney test where normal distribution could not be confirmed (left graph; p = 0.8851, 0.1804 and 0.0039 for stage 0, 1 and 2, respectively) and a paired t-test or Wilcoxon test where normal distribution could not be confirmed (right graph; 0.3225, 0.004 and 0.0609 for stage 0, 1 and 2, respectively). Scale bar: 25 μm in b. **, p < 0.01. See source data for individual data points and further statistical parameters. Illustration created with BioRender.
