Fig 5

(A, B) Quantification of cleaved caspase 3 positive cells from 48 hpf wild-type and Spint1a-deficient larvae treated for 24 hours with 10 μM FK-866 or 100 μM olaparib. (C) Representative merge images of maximum intensity projection of an apotome Z stack from zebrafish larvae of every group are shown. WIHC with anti-cleaved Casp3 (green) and anti-P63 (basal keratinocyte marker, red) were counterstained with DAPI (blue). (D–N) Pharmacological and genetic inhibition experimental settings (D, G, J). Quantification of the percentage of neutrophils out of the CHT in embryos treated with 10 nM NP (Aifm1 translocation inhibitor) (E), aifm1 genetic inhibition (H) and parga mRNA overexpression (K). Representative merge images (brightfield and red channels) of lyz:dsRED zebrafish larvae of every group are shown (F, I, L). For mRNA overexpression, 1-cell stage zebrafish eggs were microinjected, and imaging was performed in 3dpf larvae (J). Western blots with anti-PAR and anti-β-actin of tail fold lysates from 3 dpf wild-type and Spint1a-deficient zebrafish larvae microinjected with parga or GFP mRNA. The relative abundance of PAR with respect to β-actin is shown in each lane (M). Relative mRNA levels of 3 dpf wild-type zebrafish larvae microinjected with parga or GFP mRNA (N). Each dot represents one individual, and the mean ± SEM for each group is also shown. p-Values were calculated using 1-way ANOVA and Tukey multiple range test and t test. ****p ≤ 0.0001. The data underlying this figure can be found in S1 Data. ANOVA, analysis of variance; CHT, caudal hematopoietic tissue; NP, N-phenylmaleimide.