Inhibition of Fatty Acid Metabolism Ameliorates Disease Activity in an Animal Model Of

Shriver, L.P. and Manchester, M.

Inhibition of Fatty Acid Metabolism Ameliorates Disease Activity in an Animal Model of Multiple Sclerosis

Supplementary Figures and Movies

Supplementary Figure 1. Etomoxir has No effect on Body Weight in EAE and Does Not Induce Symptoms in Control mice. a) Average disease course (n=5) after MOG 35-55 induced EAE in untreated mice (). Mice developed initial disease symptoms at approximately day 10 after immunization and progressed to hind-limb motor dysfunction by day 16. CFA controls (n=4) developed no disease (). Data is represented as mean + SEM. b) Average disease score for control CFA-immunized mice treated with 15 mg/kg etomoxir dosed at days 8 and 15 (arrows, closed circles; ). c) Average weight of CFA-immunized mice receiving 15 mg/kg of etomoxir () or vehicle (). Data represents mean + SEM for 4 mice/group; neither group of mice displayed any evidence of clinical disease.

Supplementary Figure 2. Enlarged images of spinal cords from Figure 3 showing T cells (red) and merged images from a vehicle-treated mouse (a,b) and an etomoxir-treated mouse (c,d). e) Quantitation of immunohistochemistry for CD4+ T cells and CD11b+ macrophages and microglial cells in vehicle-treated (grey bars) and etomoxir-treated (black bars). Data is expressed as lesion area/total spinal cord area (mean + S.D.) and ** p < 0.01.

Supplementary Figure 3. IL-17 production in MOG-stimulated T cells. Splenocytes from mice immunized with MOG peptidewere cultured under high (black bars) or low glucose (grey bars) conditions with antigen and IL-12 in the presence of etomoxiror vehicle. Small amounts of IL-17 are detected in high glucose conditions with no difference between etomoxir and vehicle treated groups (p = 0.36). Under low glucose conditions, no IL-17 was detected in these cultures.

Supplementary Movie 1. Vehicle-treated mouse on day 20 after EAE induction. Mouse received two intraperitoneal injections of sterile water starting on day 8. Mouse displays right hind-limb paralysis typical of MOG35-55 induced EAE. Disease score is 2.

Supplementary Movie 2. Vehicle-treated mouse on day 15 after EAE induction. Mouse shows hind-limb paralysis and loss of tail tone. Disease score is 2.5

Supplementary Movie 3 . Etomoxir-treated mouse on day 20 after EAE induction. Mouse was treated with 15mg/kg etomoxir starting on day 8. A slight loss of tail tone is evident, however gross motor movement is maintained after treatment. Disease score is 1

Supplementary Movie 4. EAE mouse treated with 15 mg/kg etomoxir beginning on day 8 and observed on day 15 after EAE induction. Mouse displays loss of tail tone, but maintains hind-limb motor movement. Disease score is 1.

Supplementary Movie 5. Etomoxir-treated mouse on day 15 after EAE induction. Mouse shows tail tone and normal hind-limb movements. Disease score is 0.

1