Fig. 5

PF4 induce NETs generation through autophagy. a–c Neutrophils from healthy people and CVST patients were stained with LC3B (red), MPO (green). Control neutrophils were incubated with PRP from healthy subjects and CVST patients in the presence of an anti-PF4 antibody. d Neutrophils underwent autophagy and were defined as Cyto ID⁺ cells by flow cytometry. e–j Neutrophils from each group were co-stained with LC3B (red) and MPO (green) and observed by confocal microscopy. Neutrophils were incubated with PRP from each group and HCQ (autophagy inhibitor). k The rate of citH3 + PMN from each group was measured by flow cytometry. Statistics: Brown-Forsythe and Welch’s ANOVA test (D); Kruskal–Wallis test (k); The inset scale bars are 20 μm in a–c and e–j. ****P < 0.0001