Deficiency in TRAM hindered neutrophil elastase secretion and the blockage of elastase restored markers of endothelial integrity. a The secretion of neutrophil elastase by WT and TRAM KO neutrophils upon 24-h fMLP stimulation was evaluated by ELISA. In a separate experiment, endothelial cells were cultured in the neutrophil-conditioned media in the presence of LPA. The neutrophil-conditioned medium was collected from fMLP-stimulated WT neutrophil cultures with or without 1 μg/ml neutrophil elastase inhibitor, sivelestat. b Flow cytometry analysis of the expression of CD31, ESAM, JAM-1, and VE-Cadherin on endothelial cells cultured in two different conditioned media. ****P < 0.0001, ***P < 0.001, **P < 0.01, *P < 0.05 using two-sided Student’s t-test. n = 4. neu neutrophils. inh. inhibitor (sivelestat)