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ZFIN ID: ZDB-FIG-101011-48
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. 3

Pou5f1 and Cebpβ Directly Regulate aid, mbd4, and gadd45α Expression in apcmcr Embryos

(A) Whole-mount in situ staining for aldh1a2 in apcmcr and apcwt at 72 hpf.

(B and C) Whole-mount in situ staining for fabp2 in apcmcr and apcwt embryos (B) or in control morphant and rdh1l morphant embryos (C) (at 72 hpf) treated with vehicle or RAL (2 μM).

(D) RT-PCR for Pou5f1 (Oct4) and Cebpβ in apcmcr and apcwt treated with DMSO or ATRA (1 μM). The y axis shows fold induction normalized to 28S and wild-type DMSO-treated sample.

(E and F) Whole-mount in situ staining for aid, gadd45α, and mbd4 in apcmcr and apcwt injected with control, Pou2f1a, pou5f1, or cebpβ Mo (E) or in wild-type embryos injected with control, Pou2f1, pou5f1, or cebpβ expression plasmid (F).

(G) Graph showing fold enrichment near the aid or gadd45α TSS (containing overlapping Oct and Cebp binding sites) for Cebpβ and Pou5f1 in embryos injected with V5-Cebpβ- (along with Pou5f1 Mo, 80 pg), V5-Pou5f1-, or FLAG-Pou2f1-expressing plasmids. ChIP was performed with antibodies against the tags. Normalization control primers are located not, ~3 kb upstream (region without Cebpβ sites) of the TSS of the Gadd45α gene.

Error bars indicate ± SD. See also Figure S3.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
aicda WT control Protruding-mouth head ISH
apchu745/hu745 control Protruding-mouth head ISH
Protruding-mouth intestine ISH
apchu745/hu745 + MO1-cebpb standard conditions Protruding-mouth head ISH
apchu745/hu745 + MO1-pou2f1a standard conditions Protruding-mouth head ISH
Protruding-mouth intestine ISH
apchu745/hu745 + MO1-pou5f3 standard conditions Protruding-mouth head ISH
aldh1a2 WT standard conditions Protruding-mouth intestine ISH
apchu745/hu745 standard conditions Protruding-mouth intestine ISH
fabp2 WT control Protruding-mouth intestine ISH
apchu745/hu745 control Protruding-mouth whole organism ISH
gadd45ab WT control Protruding-mouth whole organism ISH
apchu745/hu745 control Protruding-mouth head ISH
Protruding-mouth intestine ISH
apchu745/hu745 + MO1-cebpb standard conditions Protruding-mouth head ISH
apchu745/hu745 + MO1-pou2f1a standard conditions Protruding-mouth head ISH
Protruding-mouth intestine ISH
apchu745/hu745 + MO1-pou5f3 standard conditions Protruding-mouth intestine ISH
mbd4 WT control Protruding-mouth trunk ISH
apchu745/hu745 control Protruding-mouth head ISH
Protruding-mouth trunk ISH
apchu745/hu745 + MO1-cebpb standard conditions Protruding-mouth head ISH
apchu745/hu745 + MO1-pou2f1a standard conditions Protruding-mouth head ISH
Protruding-mouth trunk ISH
apchu745/hu745 + MO1-pou5f3 standard conditions Protruding-mouth head ISH
Protruding-mouth trunk 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
apchu745/hu745 standard conditions Protruding-mouth digestive tract mesoderm development disrupted, abnormal
Protruding-mouth intestine immature, abnormal
apchu745/hu745 + MO1-cebpb standard conditions Protruding-mouth digestive tract mesoderm development temporally extended, abnormal
apchu745/hu745 + MO1-pou2f1a standard conditions Protruding-mouth digestive tract mesoderm development disrupted, abnormal
Protruding-mouth intestine immature, abnormal
apchu745/hu745 + MO1-pou5f3 standard conditions Protruding-mouth digestive tract mesoderm development temporally extended, 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