Fig. 1: Early and pervasive axonal ATP deficits in EAE lesions.
a, Experimental design for axonal ATP/ADP ratio ([ATP/ADP]axon) measurements in EAE. b, Maximum intensity projections of in vivo multi-photon image stacks of spinal cord axons of control (left) and acute EAE (right) in Thy1-PercevalHR mice. Grayscale look-up table (LUT; λex 950 nm) (top). Ratiometric [ATP/ADP]axon LUT (λex ratio 950 nm:840 nm) (bottom). L, low; H, high. c, Details from b. [ATP/ADP]axon images of control axon in healthy spinal cord and normal-appearing, swollen and fragmented axons in acute EAE (top to bottom). d, Spinal cord axons during chronic EAE shown as in b. e, [ATP/ADP]axon of single axons in healthy and EAE mice normalized to mean of controls (magenta: axons after CCCP, 100 μM; mean ± s.e.m.; 246 axons from six control, 272 axons from four EAE, acute and 522 axons from six EAE, chronic mice; compared by Kruskal–Wallis and Dunn’s multiple comparison test; values ≥ 1.5 are lined up on the ‘≥1.5’ dashed line). f, [ATP/ADP]axon gradient in a dorsal root axon traced through a lesion using in vivo multi-photon imaging. Traced axon pseudo-colored yellow, running from root at top left to the lesion center at lower right, superimposed on full volume projection, grayscale LUT, λex 950 nm (left). Boxes indicate locations of high-resolution stacks. Details showing the [ATP/ADP]axon gradient between locations far from (1) versus close to the lesion (3) (right). LUTs as in b. g, Paired analysis of [ATP/ADP]axon far versus close to the lesion (n = 22 axons from three EAE mice; two-tailed, paired t-test; normalized to control). Scale bars, 25 μm in b, also applied to d; 10 μm in c and f (right); 100 μm in f (left). ****P < 0.001. See source data for individual data points and further statistical parameters. Illustration created with BioRender.
