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. 2016 Oct 18;6:35559. doi: 10.1038/srep35559

Figure 4. Induction of NETosis in human neutrophils incubated with whole Wolbachia.

Figure 4

Representative micrographs of resting human neutrophil cultures or stimulated for 2 h with PMA, cell supernatant containing Wolbachia at an estimated ratio of 25 bacteria per neutrophil or matching concentration of Wolbachia free supernatant, subsequently stained with DAPI (white) (A) Scale bar 100 μm. Quantification of extracellular DNA stripped from cultures following stimulations with Wolbachia containing or Wolbachia free supernatant (B). Quantities of extracellular DNA released following Wolbachia-free and Wolbachia-containing cell supernatant stimulation of human neutrophils. Bars represent means (+SEM) of 6–9 replicates (2 for unstimulated Ne and Ne + PMA controls) pooled from 2 individual experiments. Matching C6/36wAlbB cell sup volumes were measured for DNA content by picoGreen DNA assay in duplicate. Difference between quantities of extracellular DNA released following Wolbachia-free and Wolbachia-containing cell supernatant stimulation of human neutrophils was evaluated by Mann-Whitney test. Significance indicated *p < 0.05.