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ZFIN ID: ZDB-FIG-101011-46
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:
Anatomical Terms:
Stage: Protruding-mouth
PHENOTYPE:
Fish:
Observed In:
Stage: Protruding-mouth

Fig. 1 Upregulation of DNA Demethylase Components, and Hypomethylated Genes in apc Mutant Embryos and FAP Adenomas

(A–D) Whole-mount in situ staining for aid (A), apobec2a (A), apobec2b (A), mbd4 (B), tdg (B), gadd45α (C), gadd45βlike (C), gadd45β (C), gadd45γ (C), and dnmt1 (D) in apc mutants (apcmcr) and siblings (apcwt) zebrafish embryos at 72 hpf.

(E) MeDIP-qPCR for genes shown (selected from genome-wide MeDIP-ChIP microarray analysis) in apcmcr and apcwt (72 hpf) that are either uninjected or injected with aaa Mo (combination of aid, apobec2a, and apobec2b morpholinos; 0.5 ng each) or mbd4 and tdg morpholinos together (1 ng each) or V5-Dnmt1 expressing plasmid (1 pg, an amount that rescues Dnmt1 morphants with 97% knockdown of Dnmt1 levels). Dnmt1 protein levels are in Figure S1C.

(F) MeDIP-qPCR for genes shown (selected from genome-wide MeDIP-ChIP microarray analysis) in human adenomas (A) and matching uninvolved tissues (U) from FAP patients. P1-P10 refers to ten different patients.

In both (E) and (F), the y axis shows values for each promoter region normalized to a negative control region lacking CpGs and then normalized to the values from wild-type or uninvolved, valued at 1.

Error bars indicate ± standard deviation (SD). See also Figure S1.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
aicda WT standard conditions Protruding-mouth head ISH
apchu745/hu745 standard conditions Protruding-mouth head ISH
Protruding-mouth intestine ISH
apobec2a WT standard conditions Protruding-mouth whole organism ISH
apchu745/hu745 standard conditions Protruding-mouth whole organism ISH
apobec2b WT standard conditions Protruding-mouth whole organism ISH
apchu745/hu745 standard conditions Protruding-mouth whole organism ISH
dnmt1 WT standard conditions Protruding-mouth whole organism ISH
apchu745/hu745 standard conditions Protruding-mouth whole organism ISH
gadd45aa WT standard conditions Protruding-mouth head ISH
Protruding-mouth intestine ISH
apchu745/hu745 standard conditions Protruding-mouth head ISH
Protruding-mouth intestine ISH
gadd45ab WT standard conditions Protruding-mouth whole organism ISH
apchu745/hu745 standard conditions Protruding-mouth whole organism ISH
gadd45ba WT standard conditions Protruding-mouth whole organism ISH
apchu745/hu745 standard conditions Protruding-mouth whole organism ISH
gadd45ga WT standard conditions Protruding-mouth whole organism ISH
apchu745/hu745 standard conditions Protruding-mouth whole organism ISH
mbd4 WT standard conditions Protruding-mouth whole organism ISH
apchu745/hu745 standard conditions Protruding-mouth whole organism ISH
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
apchu745/hu745 standard conditions Protruding-mouth digestive tract mesoderm development disrupted, abnormal
Protruding-mouth intestine immature, 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