Injury and infection induced tnfa:GFP is cyclooxygenase independent. (A) Confocal micrographs of the TgBAC(tnfα:GFP)pd1028 line at 3 days post-fertilization larvae with or without tailfin wound (induced 16 h previously) in phenol red (PR) or dominant negative (DN) Hif-1α embryos. The dotted line indicates the border of the tailfin wound. (B) Corrected fluorescence intensity levels of tnfa:GFP in larvae in (A). Mean ± SEM, n = 60 cells from 10 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. (C) Confocal micrographs of 1 dpi caudal vein region of larvae in the TgBAC(tnfa:GFP)pd1028 line. Phenol red (PR) and dominant negative Hif-1α (DN1) injected larvae were infected with Mm Crimson or PVP controls. (D) Corrected fluorescence intensity levels of tnfa:GFP in larvae in (C). 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. (E) Confocal micrographs of the TgBAC(tnfα:GFP)pd1028 line at 3 days post fertilization larvae with or without tailfin wound (induced 16 h previously) in phenol red (PR) or dominant active Hif-1α embryos. Embryos were treated with DMSO or NS398 (Cox-2 inhibitor). (F) Corrected fluorescence intensity levels of tnfa:GFP in larvae in (E). Mean ± SEM, n = 24 (in not wounded) or n = 36 (in wounded) cells from six embryos accumulated from two independent experiments. n.b. Not-wounded fish had fewer macrophages at the tailfin site. P-values shown are: *P < 0.05, **P < 0.01, and ***P < 0.001, two way ANOVA with Bonferonni post-test adjustment. (G) Confocal micrographs of 1 dpi caudal vein region of Mm Crimson infected larvae in the TgBAC(tnfa:GFP)pd1028 line. Phenol red (PR) and dominant active Hif-1α (DA1) injected larvae were treated with DMSO, SC560 (Cox-1 inhibitor), and NS398 (Cox-2 inhibitor) and infected with Mm Crimson. (H) Corrected fluorescence intensity levels of tnfa:GFP in larvae in (G). Mean ± SEM, n = 36 cells from six embryos representative of 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.
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