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ZFIN ID: ZDB-FIG-101011-50
Rai et al., 2010 - DNA demethylase activity maintains intestinal cells in an undifferentiated state following loss of APC. Cell   142(6):930-942 Full text @ Cell
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Fish:
Knockdown Reagents:
Anatomical Terms:
Stage: Protruding-mouth
PHENOTYPE:
Fish:
Knockdown Reagents:
Observed In:
Stage: Protruding-mouth

Fig. 5

Lef1 and Groucho2 Suppress rdh1l in apcmcr Zebrafish

(A) Whole-mount in situ staining for lef1, groucho2, tcf4, or groucho3 in apcwt and apcmcr embryos at 72 hpf. Red arrow points to the intestine.

(B) Whole-mount in situ staining for fabp2, trypsin, rdh1l, and irbp in apcmcr/control Mo, apcmcr /lef1 Mo, and apcmcr /groucho2 Mo embryos at 72 hpf. Groucho2 mo efficiency is in Figure S5G.

(C and D) ChIP for Lef1 on rdh1l promoter in apcwt and apcmcr embryos.

(C) Schematic showing the Lef1 sites on the rdh1l promoter and location of the primer set P1 (containing Lef1 sites) and P2 (without Lef1 sites).

(D) Graph showing fold enrichment of Lef1 occupancy at P1/P2 on rdh1l promoter in apcmcr embryos compared to apcwt at 72 hpf. Lef1 values were normalized to ones obtained with a nonspecific antibody and then expressed as fold enrichment compared to apcwt. Error bars indicate ± SD.

(E) Whole-mount in situ staining for fabp2 and gata6 in 72 hpf embryos injected with full-length lef1 or dominant negative (deficient in β-catenin binding) lef1 mRNA alone or with groucho2 mRNA.

See also Figure S5.

Gene Expression Details
Gene Antibody Fish Conditions Stage Qualifier Anatomy Assay
dhrs9 WT control Protruding-mouth intestine ISH
apchu745/hu745 control Protruding-mouth Not Detected intestine ISH
apchu745/hu745 + MO1-tle3a standard conditions Protruding-mouth intestine ISH
apchu745/hu745 + MO2-lef1 standard conditions Protruding-mouth intestine ISH
fabp2 WT control Protruding-mouth intestine ISH
WT standard conditions Protruding-mouth intestine ISH
apchu745/hu745 control Protruding-mouth Not Detected intestine ISH
apchu745/hu745 + MO1-tle3a standard conditions Protruding-mouth intestine ISH
apchu745/hu745 + MO2-lef1 standard conditions Protruding-mouth intestine ISH
gata6 WT standard conditions Protruding-mouth intestine ISH
lef1 WT standard conditions Protruding-mouth intestine ISH
apchu745/hu745 standard conditions Protruding-mouth intestine ISH
prss1 WT control Protruding-mouth pancreas ISH
apchu745/hu745 control Protruding-mouth Not Detected pancreas ISH
apchu745/hu745 + MO1-tle3a standard conditions Protruding-mouth pancreas ISH
apchu745/hu745 + MO2-lef1 standard conditions Protruding-mouth pancreas ISH
rbp3 WT control Protruding-mouth retina ISH
apchu745/hu745 control Protruding-mouth Not Detected retina ISH
apchu745/hu745 + MO1-tle3a standard conditions Protruding-mouth Not Detected retina ISH
apchu745/hu745 + MO2-lef1 standard conditions Protruding-mouth Not Detected retina ISH
tcf4 WT standard conditions Protruding-mouth intestine ISH
apchu745/hu745 standard conditions Protruding-mouth intestine ISH
tle3a WT standard conditions Protruding-mouth Not Detected intestine ISH
apchu745/hu745 standard conditions Protruding-mouth intestine ISH
tle3b WT standard conditions Protruding-mouth intestine ISH
apchu745/hu745 standard conditions Protruding-mouth intestine ISH
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
apchu745/hu745 control Protruding-mouth digestive tract mesoderm development disrupted, abnormal
Protruding-mouth intestine immature, abnormal
Protruding-mouth retina immature, abnormal
Protruding-mouth retina development in camera-type eye disrupted, abnormal
apchu745/hu745 + MO1-tle3a standard conditions Protruding-mouth intestine morphology, normal
Protruding-mouth retina immature, abnormal
Protruding-mouth retina development in camera-type eye disrupted, abnormal
apchu745/hu745 + MO2-lef1 standard conditions Protruding-mouth intestine morphology, normal
apchu745/hu745 + MO2-lef1 standard conditions Protruding-mouth retina immature, abnormal
apchu745/hu745 + MO2-lef1 standard conditions Protruding-mouth retina development in camera-type eye disrupted, abnormal
Acknowledgments:
ZFIN wishes to thank the journal Cell for permission to reproduce figures from this article. Please note that this material may be protected by copyright.

Reprinted from Cell, 142(6), Rai, K., Sarkar, S., Broadbent, T.J., Voas, M., Grossmann, K.F., Nadauld, L.D., Dehghanizadeh, S., Hagos, F.T., Li, Y., Toth, R.K., Chidester, S., Bahr, T.M., Johnson, W.E., Sklow, B., Burt, R., Cairns, B.R., and Jones, D.A., DNA demethylase activity maintains intestinal cells in an undifferentiated state following loss of APC, 930-942, Copyright (2010) with permission from Elsevier. Full text @ Cell