Fig. 6

NLRP3 promotes ROS-dependent NET formation, regardless of PAD4, in EAE pathogenesis. A Representative scatter plots and percentages of ROS⁺ neutrophils (gated on CD11b⁺ and Ly6G⁺ cells) in the brain of EAE-induced WT and NLRP3 KO mice. B and D Total RNA was isolated from the whole brain of both groups at the disease peak, and (B) Hmox1 and (D) Pad4 expression were analyzed by quantitative real-time PCR (qRT-PCR). C Immunoblotting and densitometric analysis for PAD4 and β-actin in the brains collected from both groups at peak disease. For quantification, the target blots were normalized using β-actin. E Representative histogram and relative MFI of extracellular PAD4 on the brain-infiltrated neutrophils (gated on CD11b⁺ and Ly6G.⁺ cells) from both groups. ns, non-significant. Data are obtained from more than triplicated independent experiments and presented as mean ± SEM. *P < 0.05, ***P < 0.001