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Fig. S8
Efficacy of the macrophage and neutrophil depletion strategy. (A) The double transgenic zebrafish line [tg(mpx:egfp)/(mpeg1:mCherry-F)] was injected at the one-cell stage with the pu.1 morpholino to specifically deplete macrophages. Representative overlays of transmission and red fluorescence stereomicroscope time-lapse images showing the pu.1 morpholino effect on the macrophage population in the tail region. (Scale bars, 200 µm.) (B) Representative overlays of transmission and green fluorescence stereomicroscope images showing the absence of effect of the pu.1 morpholino on the neutrophil population in the tail region at 48 hpf. (Scale bars, 200 µm.) This resulted in a total depletion of macrophages, whereas neutrophils remained unaffected. The first macrophages start to renew at 4 dpf. (C) The double transgenic zebrafish line [tg(mpx:egfp)/(mpeg1:mCherry-F)] exhibiting green neutrophils and red macrophages was injected at the one-cell stage with the csf3r morpholino to specifically deplete neutrophils. Representative overlays of transmission and green fluorescence stereomicroscope time-lapse images show the csf3r morpholino effect in neutrophil population in whole embryo. (Scale bars, 200 µm.) This resulted in a highly reduced neutrophil population. (D) Shown are the overlays of transmission and green/red fluorescence stereomicroscope images of representative 2 dpf larvae showing that the macrophage population remained unaffected in the csf3r morphants.