Figure 3.

The effects of NETs on the diffusion of doxorubicin (DOX). Neutrophils (1 × 10⁷/4 ml) stimulated with PMA (A) or LPS (B) were placed in the bottom chamber as described in Figure 2 legend and DOX added at a final concentration of 10 μM and incubated for another 30 min. In some wells, DNAse I was added at a final concentration of 1000 u/ml at 30 min before the addition of DOX. Then, culture inserts containing 2 mL of HBSS were placed in the bottom chambers and auto fluorescence intensities of DOX in the upper chamber measured at the indicated time points. In each set of experiments, the relative ratios of fluorescein intensities were calculated against the value of the samples measured at 1 h after incubation in control wells which did not contain NETs and DNAse I. Data are shown as mean ± standard deviation in 3 (PMA) and 3 (LPS) different experiments. ∗: p < 0.05, ∗∗: p < 0.01.