ZFIN ID: ZDB-FIG-150422-4
Marjoram et al., 2015 - Epigenetic control of intestinal barrier function and inflammation in zebrafish. Proceedings of the National Academy of Sciences of the United States of America   112(9):2770-5 Full text @ Proc. Natl. Acad. Sci. USA
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Gene:
Fish:
Knockdown Reagent:
Anatomical Term:
Stage: Day 4
PHENOTYPE:
Fish:
Knockdown Reagent:
Observed In:
Stage: Day 4

Fig. 4

tnfa knockdown rescues the IBD-like phenotype in the uhrf1pd1092 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 uhrf1pd1092 mutant (Upper) and tnfa morpholino-injected uhrf1pd1092 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

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
GFP uhrf1pd1092/pd1092; pd1028Tg control Day 4 intestinal epithelial cell IHC
uhrf1pd1092/pd1092; pd1028Tg + MO4-tnfa standard conditions Day 4 intestinal epithelial cell IHC
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
uhrf1pd1092/pd1092; pd1028Tg control Day 4 gut epithelium permeability, abnormal
Day 4 intestinal epithelial cell decreased height, abnormal
uhrf1pd1092/pd1092; pd1028Tg + MO4-tnfa standard conditions Day 4 gut epithelium permeability, normal
Day 4 intestinal epithelial cell decreased height, abnormal
Acknowledgments:
ZFIN wishes to thank the journal Proceedings of the National Academy of Sciences of the United States of America for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ Proc. Natl. Acad. Sci. USA