Fig. 7

BBG treatment suppressed the P2X7R/NLRP3 pathway in CD4⁺T cells in EAN. Proteins were obtained from sciatic nerves at the peak of EAN and subjected to western blot analysis and quantification. NLRP3 expression in immune cells was analyzed through flow cytometry and immunofluorescence. (A) Representative western blots of P2X7R, NLRP3, IL-1β and Caspase-1 protein expression in sciatic nerve. (B-E) Protein levels were quantified via densitometric analysis of immunoreactive bands in western blots using ImageJ software and were normalized to that of GAPDH. (F-G) Flow cytometry analysis and NLRP3 quantification on CD4⁺ T cells in splenic MNCs of control (unimmunized group), vehicle and BBG-treated rats. (H) Immunofluorescence photomicrographs of NLRP3 expression in CD4⁺T cells in the sciatic nerves of control, vehicle, and BBG-treated EAN rats. (I) Number of NLRP3-positive cells in sciatic nerve sections obtained from control rats, vehicle and BBG-treated EAN rats. (J) NLRP3 quantification on CD4⁺T cells. Scale bars, 20 μm. Experiments were performed in triplicate (n = 6 per replicate) with similar results. Control group, unimmunized group; BBG-P, preventative BBG group; BBG-T, therapeutic BBG group. *p < 0.05, **p < 0.01