Fig. 6. Transcriptional firing is required for UV-induced NETosis.

a, b DNA release following UV (1.92 J/cm²) treatment was measured using the SYTOX Green plate reader assay (n = 3). Cells were incubated with Actinomycin D (10 µM) for 1 h prior to UV treatment. Actinomycin D significantly inhibits UV-induced NETosis. c Neutrophils were incubated with Actinomycin D (10 µM) for 1 h and then treated with UV (1.92 J/cm²) and incubated for 240 min. Cells were stained for DNA (DAPI; blue). Fluorescence imaging shows that Actinomycin D inhibits UV-induced NETosis (scale bar, 10 μm). Images are representative of three independent experiments. For all the graphs: error bars represent SEM. ActD, Actinomycin D; *p < 0.05 for comparing UV treatment alone to UV treatment with Actinomycin D