Extended Data Fig. 3: Effect of NET stimulation on HBMVECs.
(a) Representative images of HBMVECs (shScramble (shScr), top and shCCM3, bottom) stimulated with 500 ng/mL NET-enriched supernatant or control (Ctrl) media for 24 h showing changes in cell morphology (n = 3). (b–g) Representative western blot images of HBMVECs (shscramble or shCCM3) stimulated with 500 ng/mL NET-enriched supernatant or control media for 24 h (n = 3 biological replicates) (B-C) and quantified (D-G; in E-G normalization was done to α-tubulin or GAPDH). (h) Representative images of HBMVECs (shScramble, top and shCCM3, bottom) stimulated with 500 ng/mL NET-enriched supernatant or control (Ctrl) media for 48 h showing showing changes in cell morphology (n = 6 biological replicates). (i–n) Representative western blot images of HBMVECs (shscramble or shCCM3) stimulated with 500 ng/mL NET-enriched supernatant or control media for 48 h (n = 6) (J) and quantified (J-N). (o) Representative images of HBMVECs (shScramble, left and shCCM3, right) stimulated with NET-enriched supernatant (bottom) or control (Ctrl; top) media for 48 h stained for VE-cadherin (red; junction) and DAPI (blue; nuclei). White arrow head highlights areas with no VE-cadherin staining. The stains were repeated independently three times with similar results. (p) Representative images of HBMVECs (shScramble, top and shCCM3, bottom) stimulated with 500 ng/mL NET-enriched supernatant or control (Ctrl) media for 72 h. The stains were repeated independently two times with similar results. In the graphs, the bar indicates the mean of each group, and the error bars represent the standard deviation. Statistical significance was determined using one-way ANOVA followed by a post-hoc Holm-sidak’s test.
