ZFIN Image: Viana et al., 2023, Fig 5

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Figure Caption

Fig 5

L. pneumophila interaction with neutrophils and neutrophil counts upon bloodstream injection of LD, HD WT GFP or HD ΔdotA L. pneumophila.

A) Neutrophil counts from control larvae (CTRL, black symbols) or Low Dose or High Dose of WT-GFP (blue or red symbols), or High Dose of ΔdotA-GFP (green symbols) injected larvae. The scheme of the zebrafish larvae has been adapted from [35] and has been previously modified from [87]. Data plotted in the same way as for macrophage counts in Fig 4. Two pooled independent experiments (n = 10 larvae scored for each condition). P < 0.05 was considered statistically significant (symbols: **** P < 0.0001; ***P < 0.001; **P < 0.01; *P < 0.05). No symbol on graphs means that not statistically differences were observed. B) Frames extracted from maximum intensity projection of in vivo time-lapse confocal fluorescence microscopy of 72hpf Tg(LysC::DsRed)^nz50 in the bloodstream (iv) with a LD, HD of WT-GFP or a HD of ΔdotA-GFP (lower panel) to follow neutrophil interaction with L. pneumophila immediately upon injection for 16 hpi. Images were taken from time lapse at different time points (0 hpi, 2 hpi, 4 hpi, 8 hpi and 16 hpi). Overlay of green (L. pneumophila) and red (neutrophils) fluorescence of the caudal area of the larvae (region boxed in the scheme on the right of the panel) is shown. Data representative of n = 12 to 16 larvae scored. Scale bar: 50μm. See also related S7 Movie. C) Details of a dying phagocytosing neutrophil, progressively rounding-up and loosing fluorescence upon HD WT injection. Arrowheads point to the dying phagocytosing neutrophils. Representatives of n = 14 larvae scored. Scale bar = 10mm. See also related S8 Movie.

Acknowledgments

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