Fig. 3

PF4 induces NET formation in CVST patients. a NPAs (CD41⁺/CD66b⁺) were examined in samples from healthy controls (n = 20) and CVST (n = 17) patients. b–e Neutrophils (from healthy people) were incubated with PRP from patients and healthy subjects and stained with CD41 (red) and MPO (green) to show the colocalization of NETs and PLTs. f Neutrophils (from CVST patients) were incubated with PRP from healthy subjects and stained with CD41 (red) and MPO (green). g Plasma PF4 levels were measured in samples from CVST patients (n = 37) and healthy subjects (n = 32). h Control neutrophils were treated with different concentrations of recombinant PF4 protein. CitH3-DNA was measured in each group (n = 10). Control neutrophils were incubated with PRP from healthy subjects and CVST patients in the presence of anti-PF4 antibodies. i NET releasing neutrophils (citH3 + Neut) was measured in cells from each group by flow cytometry. j–o Neutrophils were stained with MPO (red) and citH3 (green). Statistics: Ordinary one-way ANOVA (a, g); Brown-Forsythe and Welch’s ANOVA test (f, h, i). The inset scale bars are 20 μm in b–e, j–o. *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001. Neut, Neutrophils