Related to Fig 2.

Genetic and pharmacological inhibition of Nampt alleviates skin inflammation and restores epithelial integrity in Spint1a-deficient larvae. (A) Neutrophil distribution of wild-type and Spint1a-deficient larvae treated with the pharmacological inhibitors of Nampt GMX1778 and FK-866. (B) Representative merge images (brightfield and red channels) of lyz:dsRED zebrafish larvae of every group are shown. (C) For genetic inhibition using CRISPR/Cas-9 technology, 1-cell stage zebrafish eggs were microinjected and imaging was performed in 3 dpf larvae (D). Quantification of the percentage of neutrophils out of the CHT in Spint1a-deficient larvae upon knockdown of Nampta/Namptb. (E) Representative merge images (brightfield and red channel) of lyz:dsRED zebrafish larvae of every group are shown (F) Analysis of genome editing efficiency in larvae injected with control or nampta and namptb crRNA/Cas-9 complexes and quantification rate of NHEJ-mediated repair showing all INDELs (https://tide.nki.nl/). 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 ≤ 0.001, ****p ≤ 0.0001. (G) Inflammation and oxidative stress determination in zebrafish larvae [21]. Inflammation was scored by using 2 different approaches: (i) the lyz:dsRED zebrafish transgenic line was used to quantify the percentage of neutrophils out of the CHT, i.e., neutrophil dispersion; and (ii) the nfkb:eGFP zebrafish transgenic line was used to determine NFKB activity by quantification of fluorescence intensity in the drawn white box. For oxidative stress, analysis of fluorescence intensity of the ROI (white box) of larvae preloaded with an H₂O₂ fluorogenic probe. The data underlying this figure can be found in S1 Data. ANOVA, analysis of variance; CHT, caudal hematopoietic tissue; INDEL, insertion and deletion; Nampt, nicotinamide phosphoribosyltransferase; NHEJ, nonhomologous end joining; ROI, region of interest.

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