ZFIN Image: Ma et al., 2023, Fig. 1

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Figure Caption

Fig. 1

The 5-oxoETE pathway is induced by ear infection of zebrafish larvae with P. aeruginosa

(A) OXER1 phylogenetic tree.

(B) Relative expression levels of hcar1–3 and hcar1–4 mRNA in leukocytes compared with the rest of the body determined by FACS-sorting of dissociated neutrophil (left panel, TG(lyz:pm2-mk2)) or macrophage (right panel, TG(mpeg1: eGFP)) reporter zebrafish larvae and qRT-PCR. Error bars, SD of three different pools of zebrafish leukocytes. *p < 0.05, **p < 0.01, ***p < 0.001, n.s., p > 0.05; Student’s t test (two tailed, two sample unequal variance test).

(C) Top left panel: experimental cartoon scheme. This figure was created with BioRender. Bottom left panel: Venn diagram of zebrafish genes with ENSEMBL orthologs (ENSEMBL release 107) in human, cat, mouse, and rat. Red number, number of DIOPT-filtered NRNM genes (see STAR Methods). Right panel: sorted sGCA heatmap of significantly regulated mRNA-seq expression profiles (fold-regulation > 2, p_adj < 0.05 by Benjamini and Hochberg method) with high similarity to the respective binary expression phenotypes (correlation distance < 0.25, Figure S1B, also see STAR Methods and MATLAB sGCA script). Color scale, mRNA-seq profiles are row-scaled between 0 (blue) and 1 (pink). Shown are three biological replicates per experimental condition. Heatmap inset: significant (p_adj < 0.05) fold-upregulation of select inflammatory receptor mRNAs 2 and 4 h after PA infection as calculated by DEseq2. DIOPT-filtered NRNM genes are marked on the left side of the heatmap (zebrafish gene/human ortholog). NRNM genes with “high”-ranked (criteria see STAR Methods) human orthologs are printed in bold.

(D) Heatmap of row-scaled lipid concentrations as measured by LIPIDMAPS full panel eicosanoid mass spectroscopical analysis. Color scale, relative lipid concentrations are row-scaled between 0 (blue) and 1 (orange). Boxed numbers, absolute lipid concentrations in pmol/mg tissue (~μM). Note that the color scale does not represent these absolute lipid concentrations, but that it highlights the relative changes for each lipid. Shown are three biological replicates (i.e., ~400 3 dpf zebrafish larvae per pool) per experimental condition. Numbers right of heatmap, p values of Student’s t test (two tailed, two sample unequal variance test) comparing 0 h PA vs. 2 h PA.

See also Figure S1 and Tables S1 and S2.

Acknowledgments

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