MOG35–55 induced chronic progressive EAE model in C57BL/6 mice
10-12 weeks old female C57BL/6 mice were purchased from Shanghai Laboratory Animal Center. All animal care and experimental procedures in this study were conducted in compliance with the protocol approved by the institutional Animal Care and Use Committee at East China University of Science and Technology. Mice were housed for 7 days in a flow cabinet with a 12 h light/dark cycle and fed with standard rodent chow and tap water ad libitum. For EAE induction, mice were immunized in the flank with 100 µg MOG35-55 peptide dissolved in Complete Freund’s Adjuvant (CFA, including extract 4 mg/ml M. tuberculosis H37Ra; Beijing Biolead Biology Sci & Tech)(Najm et al., 2015). The 200 ng/mouse pertussis toxin (PTX) was injected on day 0 and day 2. Treatment with PIP or vehicle started after immunization after 12 days and kept 17 days. Clinical symptoms scoring for MOG35-55 induced EAE was evaluated as follows: score 0, no obvious changes in motor functions; score 0.5, distal paralysis of the tail; score 1, complete tail paralysis; score 1.5, mild paresis of one or both hind legs; score 2, severe paresis of hind legs; score 2.5, complete paralysis of one hind leg; score 3, complete paralysis of both hind legs.