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. 2014 Sep 3;9(9):e106813. doi: 10.1371/journal.pone.0106813

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© 2014 Ng et al

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

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Neutrophil production of HOCl was assessed using the fluorescent HOCl probe R19-S. Phagocytosis was performed by mixing neutrophils with opsonized PsA in the chloride-free buffer (0 ) for 15 min, and the cells were further incubated in the buffer containing either 0 mM or 122 for 20 min. Then, R19-S was added to incubate for an additional 15 min. Fluorescence of R19-S produced by HOCl oxidation was analyzed by flow cytometry. Mean fluorescence intensity (MFI) of each sample was obtained and compared to their 0-min time point. Statistical difference was judged by Student’s t-test. Double asterisks indicate significant differences (p<0.01, n = 4).

Inline graphic — Neutrophil production of HOCl was assessed using the fluorescent HOCl probe R19-S. Phagocytosis was performed by mixing neutrophils with opsonized PsA in the chloride-free buffer (0 ) for 15 min, and the cells were further incubated in the buffer containing either 0 mM or 122 for 20 min. Then, R19-S was added to incubate for an additional 15 min. Fluorescence of R19-S produced by HOCl oxidation was analyzed by flow cytometry. Mean fluorescence intensity (MFI) of each sample was obtained and compared to their 0-min time point. Statistical difference was judged by Student’s t-test. Double asterisks indicate significant differences (p<0.01, n = 4).