ZFIN ID: ZDB-FIG-100218-10
Stewart et al., 2009 - A histone demethylase is necessary for regeneration in zebrafish. Proceedings of the National Academy of Sciences of the United States of America   106(47):19889-19894 Full text @ Proc. Natl. Acad. Sci. USA
ADDITIONAL FIGURES
EXPRESSION / LABELING:
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Anatomical Terms:
Stage: Day 4

Fig. 3

me3K27 H3 demethylases in caudal fin. (A) Global levels of me3K27 H3 remain intact during regeneration. Immunohistochemistry of longitudinal sections of 48 hpa regenerating caudal fins. From left to right, me3K27 H3 antibody; me3K27 H3 antibody with peptide competition; msxB; PCNA; p63. The right-most panel is a non-regenerating caudal fin stained for me3K27 H3. Antibody signal (brown) and hematoxylin counterstain (blue) are shown. (B–E) Expression of putative me3K27 H3 demethylases in regenerating caudal fins. Whole mount in situ hybridization of regenerating caudal fins at 48 h post amputation (B) kdm6a.1, (C) kdm6a.2, (D) kdm6b.2, and (E) kdm6b.1. Longitudinal sections of the fins in (B–E) reveal unique expression patterns and are shown in the right panels. The epidermis (e) and mesenchyme (m) are indicated in each panel. (F) Upregulation of kdm6b.1 during regeneration. The expression of putative me3K27 H3 demethylases was examined in non-regenerating and regenerating (48 hpa) adult caudal fins by qPCR and normalized to Ribosomal Protein L18 expression. Data are expressed as average expression levels relative to non-regenerating fins from two different cohorts (12 animals each) and error bars represent the deviation from the means derived from each cohort. Similar results were obtained in several independent experiments. (G) kdm6b.1 expression in larvae caudal fin at 24 hpa [72 h post fertilization (hpf)] by in situ hybridization. (H) Kdm6b.1 demethylates me3K27 H3. myc-tagged Kdm6b.1 was immunoprecipitated from transfected 293T cells and subjected to a histone demethylase assay, followed by immunoblotting. Some panels contain blots that were stripped and re-probed with different antibodies.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
kdm6a WT physical alteration: anatomical structure Day 4 caudal fin apical epidermal cap ISH
Day 4 caudal fin regeneration epithelium ISH
kdm6al WT physical alteration: anatomical structure Day 4 caudal fin regeneration epithelium ISH
kdm6ba WT physical alteration: anatomical structure Day 4 caudal fin apical epidermal cap ISH
Day 4 caudal fin regeneration epithelium ISH
kdm6bb WT physical alteration: anatomical structure Day 4 caudal fin apical epidermal cap ISH
msx1b Ab1-msx WT physical alteration: anatomical structure Day 4 caudal fin blastema IHC
tp63 Ab1-tp63 WT physical alteration: anatomical structure Day 4 caudal fin regeneration epithelium IHC
Antibody Labeling Details
Antibody Assay Fish Conditions Stage Anatomy
Ab8-h3 IHC WT standard conditions Day 4 caudal fin
IHC WT physical alteration: anatomical structure Day 4 caudal fin blastema
IHC Day 4 caudal fin regeneration epithelium
Phenotype Details No data available
Acknowledgments:
ZFIN wishes to thank the journal Proceedings of the National Academy of Sciences of the United States of America for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ Proc. Natl. Acad. Sci. USA