Fig. 8: OPTN inhibitor SSD impairs DC function and attenuates EAE.
a Western blotting analysis of BMDCs that treated with SSD of different concentrations for 24 h before LPS stimulation. n = 3 independent experiments. b Western blotting analysis of 293T cells that treated with cycloheximide (CHX) for different time after SSD (1 μM) treatment. n = 3 independent experiments. c, d BMDCs were cultured with SSD (1 μM) for 24 h before LPS stimulation. Flow cytometry analysis for the expression of CD80, CD86, and MHC-II. n = 3 independent experiments, P(CD80) = 0.0004, P(CD86) = 0.0049, P(MHC-II) = 0.0050. e Proliferation of CFSE-labeled OT-II CD4+ T cells that incubated with BMDCs treated with or without SSD and then pulsed with OVA323–339. n = 3 independent experiments, P = 0.0009. f, g Differentiation of OT-II CD4+ T cells that incubated with BMDCs treated with or without SSD and then pulsed with OVA323-339. n = 3 independent experiments, P(IFN-γ) = 2.6E−05, P(IL-17) = 0.0028. h C57BL/6 mice (8-week-old, female) were immunized with MOG35–55 and administered daily with SSD or placebo solution intragastrically from the day of immunization. Mean clinical scores are shown. n = 8 independent animals, P(SSD-10) = 8.9E−08, P(SSD-20) = 3.1E−11, P(SSD-40) = 1.6E−13. i (Upper) Luxolfast blue (LFB) staining of spinal cords from vehicle and SSD (40 mg/kg) treated EAE mice at day 20 after immunization. Scale bars, 200 μm. (Lower) Immunofluorescent labeling of CC1 (green) in the spinal cord of indicated mice at day 20 after immunization. Scale bars, 20 μm. n = 3 independent animals, P = 0.0047. j, k Flow cytometry analysis of Th1 and Th17 cells in spleen (SP) (j) and draining lymph nodes (DLN) (k) from vehicle and SSD treated EAE mice at day 20. n = 3 independent animals, P(IFN-γ) = 0.0148, P(IL-17) = 0.0041 in (j); P(IFN-γ) = 0.0095, P(IL-17) = 0.0004 in (k). Data are presented as means ± SD. *P < 0.05; **P < 0.01; ***P < 0.001. Unpaired two-tailed Student’s t-test. Source data are provided in Source data file.
