Fig. 5.

Analysis of functional components from NET-formed medium affecting the pericyte phenotype. A Workflow for proteomics and metabolomics comparisons between NET-formed medium (PMA) and control medium (neutrophils without PMA stimulation) (proteomics, n = 3 per group; metabolomics, n = 7 per group). B Pie chart of differentially-expressed proteins and top 20 up-regulated proteins in NET-formed medium. Changes > 1.5-fold and P < 0.05 were considered significantly different. C Hierarchical clustering of differential metabolites in NET-formed medium. Differentially-expressed molecules repeatedly consistent in the same group with P < 0.05 were screened out and are highlighted in red (up-regulated) and blue (down-regulated). D RT-PCR analysis of relative CD11b mRNA expression in histone-treated MBVPs compared to controls. Cells were treated with recombinant histone peptides for 24 h. Histones are a mixture of histones 1, 2, and 3 at the a ratio of 1:1:1. E WB analysis of CD11b and ZO-1 in MBVPs incubated with histones for 48 h. Protein levels were quantified relative to GAPDH. F FACS analysis of the percentage of CD11b⁺ MBVPs incubated under specific conditions for 48 h. Data are shown as the mean ± SEM of 3 individual experiments; *P < 0.05, **P < 0.01, ***P < 0.001, ANOVA.