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. Author manuscript; available in PMC: 2019 Aug 1.
Published in final edited form as: Arterioscler Thromb Vasc Biol. 2018 Aug;38(8):1901–1912. doi: 10.1161/ATVBAHA.118.311150

Figure 1. NETs increase the endothelial expression of leukocyte adhesion molecules and augment leukocyte adhesion to ECs under flow.

Figure 1

(A–B) HSVECs were incubated with NETs (0.5 µg DNA/ml) for the indicated periods of time (left panels) or for 3 hours with various concentrations of NETs (right panels), followed by RNA extraction and determination of VCAM-1 and ICAM-1 mRNA levels by RT-qPCR. Levels of GAPDH mRNA served as an internal control for adjustment between samples. Left panels, n = 7; right panels, n = 9. P values: *<0.05, **<0.01, ***<0.001, ****<0.0001 vs. the respective controls. (C) HSVECs were incubated with NETs (0.5 µg DNA/ml) for the indicated periods of time. Whole-cell extracts were fractionated by SDS-PAGE and immunoblotted with antibodies to VCAM-1, ICAM-1, or β-actin. (D) HUVECs were treated with NETs (0.5 µg DNA/ml) for 6 hours and analyzed by immunoblot as in (C). (E) HUVECs were treated with NETs (0.5 µg DNA/ml) for 6 hours, followed by analysis of the adhesion of THP-1 cells (left panel, n = 4) or primary human monocytes (right panel, n = 3) under flow as described in the Materials and Methods section.