Fig. 6

Platelet-derived exosomes induce NET formation via HMGB1. a HMGB1 expression in exosomes was determined by western blot loaded with equal amounts of exosomal protein (6 μg/μL, 40 μg). b Exosomes were detected by HMGB1 expression using flow cytometry. c–f PMNs were cocultured with PBS-Exo or LPS-Exo, stimulated with HMGB1 (1 μg/mL), or cocultured with LPS-Exo accompanied by BoxA (10 μg/mL) for 5 h. c Western blot of p-Akt, Akt, p-mTOR, mTOR, and LC3B in PMNs. d Immunofluorescence images show LC3B expression (green) in PMNs. Nuclei were counterstained with Hoechst (blue). e NET formation was visualized by confocal microscopy with the following fluorescent dyes: Hoechst 33342 (blue) and SYTOX Green (green). f Quantification of dsDNA in the supernatant of cultured PMNs using PicoGreen fluorescent dye. All results are representative of 3 independent experiments. Graphs represent means ± SEM; *P < 0.05, **P < 0.01 compared within two groups