Fig. 4: TCA cycle enzymes, Idh3, Idh2 and Mdh2, are dysregulated in neurons in EAE.
a, Experimental design to validate dysregulated TCA cycle enzymes identified by MitoTag proteomics in EAE. b–j, Immunofluorescence analysis of Idh3a (b–d), Mdh2 (e–g) and Idh2 (h–j) in spinal cord neuronal somata (top) and axons (bottom) of control (left) or of acute EAE (right) Thy1-mitoRFP mice (TCA cycle enzymes, green; RFP, red for neuronal mitochondria in somata or neurofilament (NF) for axonal staining, respectively). Graphs (d,g,j) show area occupancy (soma, P = 0.5009 for Idh3a, 0.1371 for Mdh2 and 0.9907 for Idh2; axon, P = 0.053 for Idh3a, 0.0357 for Mdh2 and 0.5615 for Idh2), mean fluorescence intensities (MFIs) (soma, P = 0.0007 for Idh3a, 0.0003 for Mdh2 and 0.0317 for Idh2; axon, P = 0.00006 for Idh3a, 0.0357 for Mdh2 and 0.0299 for Idh2) and their product, integrated density (soma, P = 0.0046 for Idh3a, 0.0286 for Mdh2 and 0.265 for Idh2; axon, P = 0.0079 for Idh3a, 0.0023 for Mdh2 and 0.0653 for Idh2), in EAE neuronal somata and axons normalized to the mean of control (mean ± s.e.m.; compared per animal by two-tailed, unpaired Student’s t-test or Mann–Whitney U-test where normal distribution could not be confirmed from four control and four EAE mice for Idh3a and Mdh2; five control and five EAE mice for Idh2. Scale bar, 25 μm (h) applies also to b,e and their details; 10 μm (i), applies also to c and f. *P < 0.05; **P < 0.01, ***P < 0.005; ****P < 0.001; NS, not significant. See source data for individual data points and further statistical parameters. Illustration created with BioRender.
