Fig 1: LPC 18:1 optic nerve injections promote remyelination in EAE mice. A, Multivariable dimension reduction analysis (principal component analysis, left panel) and bilinear factor model analysis (partial least-squares discriminant analysis, right panel) demonstrating the separation of samples into four distinct groups. NI, No-injection control; LR, low-recovery group; HR, high-recovery group; SN, stay-normal group. B, Quantitative iTRAQ mass spectrometry analysis of EAE optic nerves injected with LPC 18:1 (no-injection control, low-recovery, high-recovery, and stay-normal groups). One-way ANOVA demonstrating 88 significant proteins. Adjusted p-value cutoff = 0.001; post hoc analysis with Fisher’s LSD. C, Correlation pattern analysis demonstrating the top 10 proteins that either increase (red) or decrease (blue) starting with the no-injection control group and progressively moving toward the SN group. Distance measure: Pearson R. D, Reactome pathway analysis of proteins that are shared between the ANOVA and correlation pattern analysis (50 proteins for positive correlation and 38 proteins for negative correlation). Reactome pathways interactors were excluded, pathway p-value = 0.05. E, Western blot analysis demonstrating a negative correlation between oligodendrocyte progenitor cell markers and mature oligodendrocyte markers (PDGFR-a: one-way ANOVA, ***p = 0.001; MBP: one-way ANOVA, ***p = 0.001; MOG: one-way ANOVA, ***p = 0.001). PDGFR-a, catalog #ab134123, Abcam; MBP: catalog #ab7349 Abcam; MOG, catalog #ab28766, Abcam. n = 3. NI, No-injection control group; LR, low-recovery group; HR, high-recovery group; SN, stay-normal group. F, Immunohistochemistry of EAE animals injected with LPC 18:1 at 150 µm demonstrating mature oligodendrocyte localization (yellow arrowhead). MBP: catalog #ab7349, Abcam; OSP: catalog #ab7474, Abcam. Blue, DAPI; green, MBP; red, OSP. G, Dose curve analysis for determining histidine dose: 10 µm was the concentration used for C13-histidine + LPC 18:1 EAE optic nerve injections. H, Mass spectrometry C13-histidine + LPC 18:1 EAE optic nerve injections demonstrating the incorporation of isobaric C13-histidine in myelin basic protein. See Extended Data Figures 3-1, 3-2, 3-3, 3-4, and 3-5.
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