Fig. 3

Inhibition of NETs alleviates allodynia in CFA mice. A. Schematic of the experimental timeline. B. Paw withdrawal threshold of mice in each group (n = 6, group: F_{1, 5} = 77.6, P = 0.0003; time: F_{4, 20} = 39.98, P < 0.0001; interaction: F_{4, 20} = 24.39, P < 0.0001). C. Paw withdrawal latency of mice in each group (n = 6, group: F_{1, 5} = 99.68, P = 0.0002; time: F_{4, 20} = 49.51, P < 0.0001; interaction: F_{4, 20} = 17.23, P < 0.0001). D–G. Representative blots and quantification of PAD4, MPO, and citH3 in the spinal cords of scrambled siRNA- and PAD4 siRNA-treated mice (n = 5). H. Immunofluorescence staining for PAD4 in the spinal cords (n = 3, scale bars: 100 μm). I. Double immunofluorescence staining for citH3 (red) and MPO (green) in the spinal cords of scrambled siRNA- and PAD4 siRNA-treated mice (n = 3). NETs visualized by colocalization of citH3 and MPO staining (merged images; n = 3, scale bars: 50 or 100 μm). J. Schematic of the experimental timeline. K. Paw withdrawal threshold of CFA mice treated with vehicle or CI-amidine (n = 6, group: F_{1, 5} = 283.9, P < 0.0001; time: F_{4, 20} = 29.81, P < 0.0001; interaction: F_{4, 20} = 11.58, P < 0.0001). L. Paw withdrawal latency of CFA mice treated with vehicle or CI-amidine (n = 6, group: F_{1, 5} = 461.2, P < 0.0001; time: F_{4, 20} = 100.9, P < 0.0001; interaction: F_{4, 20} = 41.12, P < 0.0001). M and N. Representative blots and quantification of MPO, and citH3 in the spinal cords of mice. O. Double immunofluorescence staining for citH3 (red) and MPO (green) in the spinal cords of vehicle- and CI-amidine-treated mice (n = 3). *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, compared with the scrambled siRNA or vehicle group mice