Fig. 5

Fig. 5

a Similar experiment as in Fig. 4b except Tg(lyz:TagRFP) embryos were injected with Halo-TEV-Keap1 and the indicated MO (500 μM). See also Supplementary Fig. 16. In all cases, embryos were exposed to HNE or DMF at 30 hpf, and neutrophils were counted at 36 hpf. Inset at right: Fold changes (treated/non-treated) in neutrophil counts in each knockdown background. P values in black were calculated with two-tailed unpaired Student’s t-test. P values in blue were calculated with ANOVA and Dunnett’s multiple comparisons test (see also Supplementary Fig. 1b). Note that the non-treated data sets in each plot are the same, but presented in each plot for clarity. bTg(lyz:eGFP) carmin embryos were treated at 30 hpf with DMF for 4 h, and neutrophils were counted subsequently. See also Supplementary Fig. 18a–d. Scale bars, 500 μm. Inset: analysis of fold change (DMF/non-treated) (see Fig. 2d) in neutrophil count. P values were calculated with two-tailed unpaired Student’s t-test (Supplementary Fig. 1b). c g(lyz:TagRFP) embryos were injected with MOs targeting Keap1a, Keap1b, both, or control MO (500 μM). See also Supplementary Fig. 19. At 30 hpf, embryos were treated with DMF for 6 h, then neutrophils were counted. P values in black were calculated with two-tailed unpaired Student’s t-test. P values in blue were calculated with ANOVA and Dunnett’s multiple comparisons test (see also Supplementary Fig. 1b). d Model of the Keap1-modification-specific apoptotic axis necessary for DMF mode-of-action in immune-cell depletion. Upon Keap1-electrophile-modification, Wdr1 is released from Keap1 binding. In concert with cofilin, Wdr1 promotes Bax-dependent mitochondrial-targeted activation of Caspase-3. All data present mean ± SEM. All sample sizes are listed in Supplementary Methods. Source data are provided as a Source data file.