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Fig. S1

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ZDB-FIG-090109-26

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Rai et al., 2008 - DNA demethylation in zebrafish involves the coupling of a deaminase, a glycosylase, and gadd45

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Fig. S1

Methylated plasmid induces demethylation in zebrafish
(A) pCMV-Luc was in vitro-methylated using HpaII methylase which converts CCGG sites to C^meCGG sites. Full methylation was verified by HpaII resistance and MspI susceptibility. Unmethylated (U) and methylated (Me) plasmids were subjected to southern blotting using a 736 bp probe against the luciferase gene cDNA coded on the plasmid (same as M-DNA; see Figure 1). (B) Fertilized zebrafish embryos were injected with 150 pg of in vitro-methylated pCMV-Luc (prepared as in panel A), and genomic DNA was isolated at indicated time points. Genome-wide methylation status was verified by the same procedure, with ethidium bromide staining revealing cleavage, and on the plasmid itself by Southern blotting using the same probe as in panel A. Note that both genome-wide and plasmid demethylation (steady state measurement) is first detected at ∼8 hpf and peaks at 13 hpf, whereas remethylation occurs by 28 hpf.

Expression Data

Expression Detail

Antibody Labeling

Phenotype Data

Phenotype Detail

Acknowledgments

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Reprinted from Cell, 135(7), Rai, K., Huggins, I.J., James, S.R., Karpf, A.R., Jones, D.A., and Cairns, B.R., DNA demethylation in zebrafish involves the coupling of a deaminase, a glycosylase, and gadd45, 1201-1212, Copyright (2008) with permission from Elsevier. Full text @ Cell