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ZFIN ID: ZDB-FIG-101011-47
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:
Condition:
Knockdown Reagent:
Anatomical Terms:
Stage: Protruding-mouth
PHENOTYPE:
Fish:
Knockdown Reagent:
Observed In:
Stage: Protruding-mouth

Fig. 2

APC Negatively Regulates Demethylase Expression via Retinoic Acid

(A) Whole-mount in situ staining for aid, gadd457alpha;, and mbd4 in apcmcr and apcwt embryos treated with DMSO, all-trans retinoic acid (ATRA, 1 μM), or Cox-2 inhibitor NS-398 (10 μM) or injected with control morpholino or ctbp1 morpholino (2 ng). β-catenin knockdown efficiency with NS-398 treatment is depicted in Figure S2A.

(B and C) Quantitative RT-PCR for aid, mbd4, and gadd45α in apcmcr and apcwt embryos treated with DMSO or ATRA (B) or in different RA deficiency models in zebrafish, as shown in (C). Expression of genes are first normalized to 28S and then to the control embryo mRNA/28S ratio, valued as 1.

(D) MeDIP-qPCR for genes shown (as in Figure 1E) in apc mutants (72 hpf) that are either untreated or treated with ATRA.

Error bars indicate ± SD. See also Figure S2.

Gene Expression Details
Gene Antibody Fish Conditions Stage Qualifier Anatomy Assay
aicda WT control Protruding-mouth head ISH
apchu745/hu745 control Protruding-mouth head ISH
Protruding-mouth intestine ISH
apchu745/hu745 chemical treatment: all-trans-retinoic acid Protruding-mouth head ISH
Protruding-mouth intestine ISH
apchu745/hu745 + MO1-ctbp1 standard conditions Protruding-mouth head ISH
Protruding-mouth intestine ISH
gadd45ab WT control Protruding-mouth Not Detected whole organism ISH
apchu745/hu745 control Protruding-mouth head ISH
Protruding-mouth intestine ISH
apchu745/hu745 chemical treatment: all-trans-retinoic acid Protruding-mouth intestine ISH
apchu745/hu745 + MO1-ctbp1 standard conditions Protruding-mouth intestine ISH
mbd4 WT control Protruding-mouth Not Detected whole organism ISH
apchu745/hu745 control Protruding-mouth trunk ISH
apchu745/hu745 chemical treatment: all-trans-retinoic acid Protruding-mouth trunk ISH
apchu745/hu745 + MO1-ctbp1 standard conditions Protruding-mouth trunk ISH
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
apchu745/hu745 control Protruding-mouth DNA demethylation increased occurrence, abnormal
apchu745/hu745 control Protruding-mouth DNA hypomethylation chronic, abnormal
apchu745/hu745 control Protruding-mouth intestine immature, abnormal
apchu745/hu745 control Protruding-mouth retinoic acid receptor signaling pathway disrupted, abnormal
apchu745/hu745 + MO1-ctbp1 standard conditions Protruding-mouth retinoic acid receptor signaling pathway 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