EAE induction and scoring
Tslpr-/- mice and WT mice were bred under
specific pathogen-free (SPF) conditions.Tslpr-/- mice were purchased from Shanghai
Biomodel Organism Science & Technology Development Co.,Ltd. FemaleTslpr-/- and WT mice (10-12 weeks old) were
immunized subcutaneously (s.c.) with MOG35-55 (Beyotime,
China) with 4 mg/ml heat-inactivated Mycobacterium tuberculosis H37Ra
(BD) at day 1 to induce EAE as described before 29.
200 ng/mouse pertussis toxin (List Biological Laboratories Inc.) was
injected i.p. at day 0 and day 2. Mice were sacrificed at day 15 and
brain tissue was collected for western blot or immunohistochemistry.
Symptoms of EAE paralysis in mice was scored as follows: 0, no disease;
1, tail weakness; 2, paraparesis; 3, paraplegia; 4, paraplegia with
forelimb weakness; 5, moribund or dead animals. To evaluate the
contribution of NLRP3 inflammasome or JAK to CNS inflammation, i.p.
injection of 50 mg/kg MCC950 (dissolved in DMSO) 30 or
oral administration of ruxolitinib (90 mg/kg/d) 31after MOG35-55 immunization were performed.