Fig. 6: Inhibition of BCAT1 ameliorates EAE severity.

Experimental autoimmune encephalomyelitis (EAE) was induced by MOG_35–55 in a CFA emulsion with PTX. Bi2 (10 mg/kg) was intraperitoneally administered to MOG-immunized mice 4 h before immunization, and the treatment was repeated three times per week for 14 days. a Clinical scores of EAE mice. b Histological analysis of spinal cord tissue stained with Luxol fast blue or Hematoxylin & Eosin on Day 16. The areas of demyelination (left) and inflammatory cell infiltration (right) are marked with dashed black lines. c Quantification of immune cell infiltration in the spinal cord (SCMCs: spinal cord mononuclear cells) among the four different groups. d The proportion of CD3⁺CD4 ⁺ T cells among total CD45⁺ T cells in untreated or Bi2-treated EAE mice (n = 6 per group). e, f Flow cytometric analysis of IL-17A- and IFN-γ-producing CD3⁺CD4⁺ T cells in the SCMCs and inguinal lymph nodes (iLNs) of EAE mice (n = 5–8). g Cell lysates were prepared from SCMCs from DMSO- or Bi2-treated EAE mice and immunoblotted for HIF-α (n = 5 per group). h A representative histogram of intracellular HIF-1α in total CD45⁺ SCMCs (left) and CD3⁺CD4⁺ T cells (right) from EAE mice (n = 5 per group). The graphs show the means ± SEMs. *p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001 by two-way ANOVA (a) or the Mann‒Whitney U test (c–h).