Fig. 4.

Effects of NET-formed medium on pericytes. MBVPs were incubated with specific media for 48 h [none, normal medium; control, medium from non-stimulated neutrophils; PMA, NET-formed medium stimulated by PMA (100 ng/mL); PMA+Cl-Amidine, medium from PMA stimulation (100 ng/mL) combined with NET inhibitor Cl-Amidine (10 μmol/L)]. Supernatant was collected by centrifugation for removing neutrophils. A Cytometric analysis of CD11b⁺ MBVPs after different treatments. B Percentage of CD11b⁺ MBVPs in A (n = 3). C Immunostaining of tight junction protein (ZO-1, green) and pericyte marker (PDGFRβ, red) on MBVPs (scale bar, 20 μm). D WB of CD11b and ZO-1 expression in MBVPs after different treatments (protein levels relative to GAPDH loading control). E RT-PCR analysis of relative mRNA levels of CD11b in different treatment groups compared to the none group (5 individual experiments per group). F Representative graph of continuous TEER values of MBVPs incubated with different culture media. TEER values of each group were compared to blank (MBVPs with normal culture medium). Data are shown as the mean ± SEM; *P < 0.05, **P < 0.01, ***P < 0.001, ANOVA.