ZFIN ID: ZDB-FIG-180827-52
Zhao et al., 2018 - Deficiency in class III PI3-kinase confers postnatal lethality with IBD-like features in zebrafish. Nature communications   9:2639 Full text @ Nat. Commun.
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Antibodies:
Fish:
Anatomical Terms:
Stage: Days 7-13
PHENOTYPE:
Fish:
Observed In:
Stage: Days 7-13

Fig. 6

PIK3C3 deficiency induces polarity defects in IECs. a Immunofluorescence staining of gut cross sections at 8 dpf. In WT embryos, columnar IECs are arranged into a sheet and E-Cadherin are laterally distributed. IECs become disorganized and E-Cadherin is endocytosed and forms intracellular aggregates in the mutants. LAMP1 is a lysosome marker and it does not co-localize with E-Cadherin aggregates. Other polarity-related proteins including Na,K-ATPase alpha 1 subunit (ATP1A1) and tight junction protein ZO-1 also lose their polarized distributions in the mutant guts. Scale bar, 20 µm. b Western blot analysis for the expression levels of E-Cadherin and ZO-1 in the dissected digestive tracts from 8 dpf embryos. Beta actin is the loading control. c qRT-PCR analysis of cdh1 and zo-1. Assays are performed as described in Fig. 3f. d Knockdown efficiency of PIK3C3 by siRNA in Caco2 cells as determined by qRT-PCR. ACTB is the internal control and data represent mean ± SD from three biological repeats (***p < 0.001 by unpaired two-tailed Student’s t-test). e PIK3C3 knockdown induces internalization of E-Cadherin in Caco2 cyst cultured on 3D Matrigel. Scale bar, 10 µm. f Quantification of (e). In each experiment, about 100 cysts were analyzed and data represent mean ± SD of the percentages of cyst with internalized E-Cadherin from three biological repeats (**p < 0.01 by unpaired two-tailed Student’s t-test). g Western blot analysis of E-Cadherin protein levels in control or siPIK3C3-treated Caco2 cells. Beta actin is the loading control

Gene Expression Details No data available
Antibody Labeling Details
Antibody Assay Fish Conditions Stage Anatomy
Ab1-actb IHC TU standard conditions Days 7-13 intestinal epithelium
IHC pik3c3zf2160/zf2160 standard conditions Days 7-13 intestinal epithelium
IHC pik3c3zf2161/zf2161 standard conditions Days 7-13 intestinal epithelium
Ab1-tjp1 IHC TU standard conditions Days 7-13 intestinal epithelial cell tight junction
IHC pik3c3zf2160/zf2160 standard conditions Days 7-13 intestinal epithelial cell
IHC pik3c3zf2161/zf2161 standard conditions Days 7-13 intestinal epithelial cell
Ab2-cdh1 IHC TU standard conditions Days 7-13 intestinal epithelial cell lateral margin
IHC pik3c3zf2160/zf2160 standard conditions Days 7-13 intestinal epithelial cell
IHC pik3c3zf2161/zf2161 standard conditions Days 7-13 intestinal epithelial cell
Ab2-lamp1 IHC TU standard conditions Days 7-13 intestinal epithelial cell lysosome
IHC pik3c3zf2160/zf2160 standard conditions Days 7-13 intestinal epithelial cell lysosome
IHC pik3c3zf2161/zf2161 standard conditions Days 7-13 intestinal epithelial cell lysosome
Phenotype Details
Fish Conditions Stage Phenotype
pik3c3zf2160/zf2160 standard conditions Days 7-13 intestinal epithelial cell ab1-tjp1 labeling spatial pattern, abnormal
Days 7-13 intestinal epithelial cell ab2-cdh1 labeling spatial pattern, abnormal
pik3c3zf2161/zf2161 standard conditions Days 7-13 intestinal epithelial cell ab1-tjp1 labeling spatial pattern, abnormal
Days 7-13 intestinal epithelial cell ab2-cdh1 labeling spatial pattern, abnormal
Acknowledgments:
This image is the copyrighted work of the attributed author or publisher, and ZFIN has permission only to display this image to its users. Additional permissions should be obtained from the applicable author or publisher of the image. Full text @ Nat. Commun.