Figure 7

CXCR3 is reduced in the absence of 12-LOX. (A) Chemokine receptor mRNA expression in macrophages from WT and Alox15–/– mice (n = 3–4 mice per genotype; *P < 0.05 by unpaired 2-tailed t test). (B) Representative contour plots showing gating of F4/80+ CXCR3+ cells are shown on the left and F4/80+ CXCR3+ cells as a percentage of total peritoneal cells from WT and Alox15–/– mice are shown on the right (n = 3 mice per genotype; *P < 0.05 by unpaired 2-tailed t test). (C) cxcr3.2 mRNA expression from whole lysates of control-treated and alox12 MO-treated zebrafish (n = 4 independent experiments from pooled zebrafish larvae; *P < 0.05 by unpaired 2-tailed t test). (D) Representative images of injured Tg(mpeg:eGFP) zebrafish tailfins stained with GFP (macrophages, green) and TO-PRO3 (nuclei, red) are shown on the left; and corresponding quantitation of relative number of macrophages at the tail injury site in the control, alox12 MO–injected fish, fish injected with a vector for macrophage-specific overexpression of cxcr3.2, and fish coinjected with alox12 MO and vector for macrophage-specific overexpression of cxcr3.2 is shown on the right (n = 20–30 fish per condition; *P < 0.05 by 1-way ANOVA with post hoc Tukey’s test). (E) Representative images of injured Tg(mpeg:eGFP) zebrafish tailfins stained with GFP (macrophages, green) and TO-PRO3 (nuclei, red) are shown on the left, and corresponding quantitation of relative number of macrophages at the tail injury site in the vehicle control and BLT2 inhibitor LY255283-treated fish is shown on the right (n = 20–25 fish per condition; *P < 0.05 by 1-way ANOVA with post hoc Tukey’s test). Scale bars in D and E indicate 100 μm. (F) Cxcr3 mRNA expression in mouse peritoneal cells treated with 10 μM LY255283 for 24 hours (cells from n = 5 mice per condition; *P < 0.05 by unpaired 2-tailed t test). (G) F4/80+CXCR3+ cells as a percentage of total peritoneal cells by flow cytometry after treatment with 10 μM LY255283 for 24 hours (cells from n = 5 mice per condition; *P < 0.05 by unpaired 2-tailed t test). Data are presented as mean ± SEM. 12-LOX, 12-lipoxygenase; MO, morpholino; GFP, green fluorescence protein; Tg(mpeg:eGFP), transgenic fish containing eGFP-labeled macrophages.