Blocking 15-lipoxygenase or leukotriene B4 receptors does not abrogate DA-Hif-1α-upregulation of tnfa:GFP. (A) Schematic of the arachidonic pathway indicating where the inhibitors act. (B) Confocal micrographs of 1 dpi caudal vein region of Mm mCrimson and PVP infected larvae in the TgBAC(tnfa:GFP)pd1028 line. Phenol red (PR) and dominant active Hif-1α (DA1) injected larvae were treated with DMSO or PD146176 (15-Lipoxygenase inhibitor). Dotted lines indicate the yolk extension of the larvae where there is non-specific fluorescence. (C) Corrected fluorescence intensity levels of tnfa:GFP in larvae in (B). Mean ± SEM, n = 42 cells from seven embryos accumulated from three independent experiments. P-values shown are: *P < 0.05, **P < 0.01, and ***P < 0.001, two way ANOVA with Bonferonni post-test adjustment. (D) Bacterial burden at 4 dpi after injection of DA Hif-1α (DA1) and 24 h of the 15-lipoxygenase inhibitor PD146176, using DMSO as a negative solvent control. Data shown are mean ± SEM, n = 48–50 as accumulated from three independent experiments. P-values shown are: *P < 0.05, **P < 0.01, and ***P < 0.001, two way ANOVA with Bonferonni post-test adjustment. (E) Confocal micrographs of 1 dpi caudal vein region of Mm mCrimson and PVP infected larvae in the TgBAC(tnfa:GFP)pd1028 line. Phenol red (PR) and dominant active Hif-1α (DA1) injected larvae were treated with DMSO or U75302/LY255283 (BLTR1/2 inhibitors). (F) Corrected fluorescence intensity levels of tnfa:GFP confocal z-stacks in uninfected larvae (PVP, empty bars) and infected larvae (Mm, filled bars) at 1 dpi of data shown in (E) after treatment with DMSO or U75302/LY255283 (BLTR1/2 inhibitors). Data shown are mean ± SEM, n = 30 cells from five embryos accumulated from two independent experiments. P-values shown are: *P < 0.05, **P < 0.01, and ***P < 0.001, two way ANOVA with Bonferonni post-test adjustment.
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