Figure 5.

Enhanced NETs release from HL-60-derived neutrophils by oxLDL treatment is associated with increased inflammatory responses in HAECs. (A) Experimental procedure for the activation of HAECs by NETs components. HL-60-derived neutrophils were stimulated with 50 nM PMA for 30 min. The culture supernatant was discarded, and the cells were washed with fresh medium to remove PMA, then the cells were subjected to subsequent treatment with 20 μg/mL LDL or oxLDL for 2 h. The culture medium of the HL-60-derived neutrophils (333 μL) was transferred to 1 mL of HAECs culture medium (final concentration of 5 μg/mL lipoproteins) and cultured for 24 h. (B) Expression of MMP-1 in HAECs was analyzed by using western blot. The intensity of each band was normalized by the corresponding band of β-actin. Quantitative data presented in the bar graph are the mean ± SD of three experiments. Asterisks indicate statistical significance against the condition PBS without PMA; ^***p < 0.001.