Fig. 4: The proinflammatory effect of FPP on PMN depends on calcium and TRP channels.

A, B The statistical results of proinflammatory cytokines (N = 4, (A) and NETs-dsDNA (N = 6, (B) in FPP-stimulated BD and HC PMN were detected by qRT-PCR and pico-green dsDNA fluorescent probe, respectively, in the presence or absence of RR and calcium. C, D Representative immunofluorescence staining (C) of 60 μg/mL FPP-induced NETosis together with RR or DNase I, as well as its quantification (D) N = 15); scale bar = 10 μm. E Quantitative analysis of RR and DNase I affecting the expression of proinflammatory cytokines and adhesion molecules in VECs (N = 5). F, G Inhibitory efficiency of RR on FPP-induced proinflammatory cytokine expression (N = 4) and NETs-dsDNA release (N = 6) in PMN of BD and HC. H 60 μg/mL FPP- and calcium-induced calcium fluxes in the PMN of BD and HC (N = 3). N for all experiments were biological replicates. Data are presented as mean ± SD; error bars indicate the SD. A two-sided p-value < 0.05 was considered statistically significant, with *p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001 indicating significant differences. Source data are provided as a Source Data file. Independent t-tests were used in (G). One-way ANOVA tests were used in (A, B and D), and two-way ANOVA tests were used in (F), with the p-value adjusted for multiple comparisons by FDR using the two-stage linear step-up procedure of Benjamini, Krieger and Yekutieli. DNase I, Deoxyribonuclease I; RR, Ruthenium Red.