FIGURE 1.

Pulmonary HMVECs display increased ICAM‐1 expression and a stronger and earlier response to LPS than HUVECs. (a) Histogram of ICAM‐1 distribution after 5 h LPS as measured with FACS. Y‐axis top to bottom represents increasing concentrations of LPS treatment of HUVEC and pulmonary HMVECs (pulmonary) as indicated. Condition 0 represents PBS CTRL treatment. X‐axis represents the increasing fluorescent intensity of ICAM‐1 measured with anti‐ICAM‐1‐FITC antibody. All cells were selected for PECAM‐1 and VE‐cadherin double positivity. (b) Quantification of ICAM‐1 surface expression after treatment of pulmonary HMVECs (pulmonary) and HUVEC with PBS (CTRL) or 10 ng/mL LPS as measured with FACS. All PBS‐treated HUVEC data points were divided by their average to indicate variation. All pulmonary HMVECs data points were divided by PBS‐treated HUVEC data points for fold difference. The dotted line indicates 1‐fold. ns = non‐significant, **p < 0.01, ***p < 0.001, n = 4. (c) Number of transmigrated neutrophils during flow TEM assay in HUVEC and HMVEC (Pulmonary) treated as indicated. Each data point represents average neutrophil TEM per field of view per condition. Three fields of view per channel were obtained during time‐lapse imaging, followed by tile scans with 25 additional fields of view per condition and replicate at the endpoint to determine the distribution of neutrophils. Bar represents an average of all experiments per condition combined. nd = not detected, **p < 0.01, error bars are presented as standard error of mean (SEM), n = 3. (d) Representative images from TEM assay. Black star indicates an area with a small cluster of transmigrated PMNs