Fig. S5

(A) Relative frequency of infected larvae demonstrating demand-driven granulopoiesis following treatment (from 6 hpi) with the pan-NOS inhibiting drug L-NMMA, relative to DMSO control treatment. (B) Relative frequency of infected larvae demonstrating demand-driven granulopoiesis following treatment (from 6, 12, 24 and 36 hpi) with the NOS2-specific inhibiting drug 1400W, relative to DMSO control treatment. Numbers represent numbers of larvae in each treatment group. (C) RT-PCR detection of disrupted nos2a splicing at 1 and 2 dpi (infected) following Nos2a MO injection. (D) Percentage survival of Control MO- and Nos2a MO-injected larvae following infection, as scored at 2 dpi (means standard ± deviations). Individual data points represent individual infection experiments (mean n=45 for Control MO and n=54 for Nos2a MO infection experiments). Infection experiments were cfu dosage-matched between Control MO and Nos2a MO cohorts. (E) Control MO- and Nos2a MO-injected Tg(lyz:EGFP)/Tg(gata1:DsRED) larvae following infection, compared to PBS controls at 2dpi. Numbers represent frequency of larvae with displayed phenotype. (F) Effect of Nos2a-depletion on steady-state granulopoiesis. Relative quantities of whole-larvae fluorescent cells, as determined by flow cytometry, within Tg(lyz:DsRED) larvae at 2, 3 and 4 dpf following injection of control and Nos2a-targeting MOs, relative to uninjected siblings. All views, anterior to left. Scale bar, 100m in E. Abbreviation: n.s., not significant.