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ZFIN ID: ZDB-FIG-101011-49
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 Range: Long-pec to Protruding-mouth
PHENOTYPE:
Fish:
Knockdown Reagents:
Observed In:
Stage Range: Long-pec to Protruding-mouth

Fig. 4

Demethylase Activity Is Required to Maintain Progenitor Cell Populations in apc Mutants

(A) Whole-mount in situ staining for aldh1a2, fabp2, and hoxa13a in apcmcr and apcwt embryos injected with control Mo, aaa Mo, combined mbd4 and tdg Mo, or overexpression of dnmt1 as in Figure 1.

(B) Cross-sections of the intestines of apcmcr and apcwt embryos whole-mount in situ stained with aid (52 hpf) or mbd4 (72 hpf). Asterisks show pronephric ducts, and arrows point to the intestine. N, notochord; SC, spinal cord.

(C) MeDIP-qPCR for genes shown in the GFP-positive cells from gutGFP;apcmcr and gutGFP;apcwt embryos (72 hpf). FACS parameters and efficiency are shown in Figures S4A and S4B.

(D–F) Immunostaining (D), RT-PCR (E), and Me-DIP PCR (F) for the indicated genes in the intestines from adult wild-type zebrafish treated with DMSO or DEAB for 7 days. In (E), RT-PCR was performed on epithelial cells isolated from the intestinal crypts.

Error bars indicate ± SD. See also Figure S4.

Gene Expression Details
Gene Antibody Fish Conditions Stage Qualifier Anatomy Assay
aicda WT standard conditions Long-pec intestine ISH
apchu745/hu745 standard conditions Long-pec intestine ISH
aldh1a2 WT control Protruding-mouth intestine ISH
apchu745/hu745 control Protruding-mouth intestine ISH
apchu745/hu745 + MO1-aicda + MO1-apobec2a + MO1-apobec2b standard conditions Protruding-mouth intestine ISH
apchu745/hu745 + MO1-mbd4 + MO1-tdg.1 standard conditions Protruding-mouth intestine ISH
fabp2 WT control Protruding-mouth intestine ISH
apchu745/hu745 control Protruding-mouth Not Detected intestine ISH
apchu745/hu745 + MO1-aicda + MO1-apobec2a + MO1-apobec2b standard conditions Protruding-mouth intestine ISH
apchu745/hu745 + MO1-mbd4 + MO1-tdg.1 standard conditions Protruding-mouth intestine ISH
hoxa13a WT control Protruding-mouth posterior intestine ISH
apchu745/hu745 control Protruding-mouth intestine ISH
apchu745/hu745 + MO1-aicda + MO1-apobec2a + MO1-apobec2b standard conditions Protruding-mouth posterior intestine ISH
apchu745/hu745 + MO1-mbd4 + MO1-tdg.1 standard conditions Protruding-mouth posterior intestine ISH
mbd4 WT standard conditions Protruding-mouth Not Detected intestine ISH
apchu745/hu745 standard conditions Protruding-mouth intestine ISH
Protruding-mouth pronephric duct ISH
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
apchu745/hu745 standard conditions Long-pec digestive tract mesoderm development disrupted, abnormal
Long-pec intestine immature, abnormal
apchu745/hu745 + MO1-aicda + MO1-apobec2a + MO1-apobec2b standard conditions Protruding-mouth intestine morphology, normal
apchu745/hu745 + MO1-mbd4 + MO1-tdg.1 standard conditions Protruding-mouth intestine morphology, normal
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