Trinitrobenzene sulfonic acid (TNBS) haptenization is concentrated to the intestinal tract and also causes extra-intestinal effects. A: Transmitted light microscopy of 6 days postfertilization (dpf) larvae exposed to increasing concentrations of TNBS. B: Confocal microscope images of whole mount immunofluorescent detection of TNBS hapenization by anti-dinitrophenyl (DNP) antibody in TG(ifabp:RFP) larvae (left: control, middle: high haptenization, right: low haptenization). C: Immunofluorescent detection of TNBS haptenization in a physical section of the intestinal bulb of a larva exposed to TNBS. Slides were counterstained with DAPI (4′,6-diamidine-2-phenylidole-dihydrochloride) to mark nuclei. D: Comparison of 5-bromo-2′-deoxyuridine (BrdU) incorporation in 6 dpf larvae following TNBS exposure. Counts were performed as described in the methods (n = 10, P values calculated by t-test, error bars represent a 95% confidence interval of the mean). E: Analysis of pro-inflammatory cytokine expression in 6 dpf larval dissected intestines and larval carcasses, measured by quantitative polymerase chain reaction (qPCR) after TNBS exposure and compared with controls. P values calculated by one-sided t-test against a hypothetic value of 1, error bars represent a 95% confidence interval of the mean, n = 3, pools of 15 larvae. *P < 0.05, **P < 0.01, ***P < 0.001.
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