Fig. 3: IRAK-M reduces gene and protein expressions of NLRP3 inflammasome, IL-1β, and GSDMD in the lumbosacral spinal cord during peak disease.

A Relative fold expression of IRAK-M mRNA in the spinal cord of AAV^IRAK-M mice compared to AAV^CTL and WT mice before EAE immunization (n = 5, respectively). ***P < 0.001 compared to the CFA group. ^###P < 0.001 compared to the AAV^CTL group. B Quantification of mRNA expression levels of NLRP3 inflammasome-related genes were assessed by RT-PCR using lumbosacral spinal cord obtained at peak disease from IRAK-M^−/− EAE and WT EAE groups (n = 6, respectively). Values represent relative fold changes compared to AAV^IRAK-M EAE mice and were normalized to GAPDH. Intravenously injected AAV with IRAK-M suppressed the CNS infiltration of NLRP3 inflammasome-related cells and ameliorated EAE. C The left panel shows representative western blotting images of indicated protein levels in lumbosacral spinal cord slices at peak disease from WT + EAE, IRAK-M^−/− + EAE, AAV^CTL + EAE, and AAV^IRAK-M + EAE mice. The right panel shows quantification analysis of western blot (normalized to GADPH) (n = 4 per group). D Immunohistochemistry was performed to detect the expression of NLRP3, ASC, caspase-1, GSDMD, and IL-1β in the lumbosacral spinal cord. The left panel shows the representative images. The right panel shows the quantification of cell numbers. Three sections per mouse were evaluated (n = 4 per group). Scale bar: 50 μm. Kruskal–Wallis plus Dunn’s test or one-way ANOVA, followed by Tukey’s post hoc analysis, was used for quantification. All data are shown as mean ± SEM. ^**P < 0.01, ^***P < 0.001.