Fig. 4

tnfa knockdown rescues the IBD-like phenotype in the uhrf1^pd1092 gut. (A) RT-PCR for tnfa from 103 hpf larvae after injection of the tnfa splice-blocking morpholino. The morpholino resulted in the inclusion of the 112-bp intron between exons 1 and 2 (top arrow; bottom arrow marks endogenous product). (B) Confocal images of cross-sections of genotyped uhrf1^pd1092 mutant (Upper) and tnfa morpholino-injected uhrf1^pd1092 mutant (Lower) larvae at 103 hpf expressing TgBAC(tnfa:GFP). (C) TgBAC(tnfa:GFP) fluorescence quantification (measured in arbitrary units, a.u.) revealed a decrease in intestinal fluorescence of genotyped mutants after morpholino injection. (D) Tg(NFκB:EGFP) fluorescence quantification (measured in arbitrary units, a.u.). Intestinal fluorescence was decreased in genotyped mutants after morpholino injection. (E) Quantification of cell shedding: Cell shedding was decreased in genotyped morpholino-injected mutants. (F) Cell height measurement: Cell height was increased in genotyped morpholino-injected mutants. (G) Dextran leakage quantification. Improved barrier function was observed in morpholino-injected mutants. (C and E: n = 9, WT; n = 16, mutant; n = 21, mutant + morpholino; D: n = 14, WT; n = 7, mutant; n = 16, mutant + morpholino; F: n = 43, WT; n = 53, mutant; n = 40, mutant + morpholino) Phalloidin, red. (Scale bars: 50 µm.) Results are shown as mean ± SEM. Significant differences between the means were detected after an ANOVA. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, n.s., not significant. Show Less