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Fig. 4

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ZDB-FIG-101011-49

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Rai et al., 2010 - DNA demethylase activity maintains intestinal cells in an undifferentiated state following loss of APC

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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 apc^mcr and apc^wt 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 apc^mcr and apc^wt 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;apc^mcr and gutGFP;apc^wt 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.

Expression Data

Genes:	aicda aldh1a2 fabp2 hoxa13a mbd4
Fish:	WT apc^hu745/hu745 apc^hu745/hu745 + MO1-mbd4 + MO1-tdg.1 apc^hu745/hu745 + MO1-aicda + MO1-apobec2a + MO1-apobec2b
Knockdown Reagents:	MO1-aicda MO1-apobec2a MO1-apobec2b MO1-mbd4 MO1-tdg.1
Anatomical Terms:	intestine posterior intestine pronephric duct
Stage Range:	Long-pec to Protruding-mouth

Expression Detail

Antibody Labeling

Phenotype Data

Fish:	apc^hu745/hu745 apc^hu745/hu745 + MO1-mbd4 + MO1-tdg.1 apc^hu745/hu745 + MO1-aicda + MO1-apobec2a + MO1-apobec2b
Knockdown Reagents:	MO1-aicda MO1-apobec2a MO1-apobec2b MO1-mbd4 MO1-tdg.1
Observed In:	digestive tract mesoderm development intestine
Stage Range:	Long-pec to Protruding-mouth

Phenotype Detail

Acknowledgments

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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