- Title
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Tet1 facilitates hypoxia tolerance by stabilizing the HIF-α proteins independent of its methylcytosine dioxygenase activity
- Authors
- Wang, J., Zhang, D., Du, J., Zhou, C., Li, Z., Liu, X., Ouyang, G., Xiao, W.
- Source
- Full text @ Nucleic Acids Res.
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Knockout of tet1 in zebrafish causes more apoptotic cells in the brain and reduction of hypoxia-inducible gene expression under hypoxia. (A) Compared with those in the brain of wild-type siblings, more apoptotic cells were detected in the brain of tet1-null zebrafish by TUNEL assays under hypoxia (5% O2, 5 h). (B) Expressions of hypoxia-inducible genes including phd3, glut1, vegfa were reduced significantly in the brain of tet1-null zebrafish embryos and adults compared with those of their wild-type siblings under hypoxia. (C) Overexpression of both zebrafish tet1 and its enzymatic inactive form (zf-tet1-mut) could activate the hypoxia response element luciferase (HRE-luc.) reporter in the N2a cell line under hypoxia. N2a cells were transfected with the indicated plasmids and luciferase activity was measured 20–24 h after transfection. The left red box indicates the alignment of partial Tet1 sequences surrounding Tet1 enzymatic activity region and the enzymatic inactive mutant (mut) with histidine mutated to tyrosine and aspartic acid mutated to alanine. zf-tet1-mut, zebrafish tet1 mutant; mu-Tet1-mut, mouse Tet1 mutant. EXPRESSION / LABELING:
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Oxygen consumption rate is not affected and DNA demethylation in brain is barely reduced when tet1 is knocked out in zebrafish. (A) Loss of tet1 in Oxygen consumption rate was the same between tet1-null zebrafish and their wild-type siblings. (B) Immunohistochemistry staining for 5hmC expression indicates that loss of tet1 caused 5hmC expression barely diminished in zebrafish brain. |
Another tet1-null zebrafish line (tet1 ihb2018/ihb2018) also exhibits less tolerant to hypoxia. (A) Tet1-null zebrafish exhibit more sensitive to hypoxia (5%) compared to their wild-type siblings. (B) (C) Expressions of hypoxia-inducible genes including phd3 and vegfa were reduced significantly in the brain of tet1-null zebrafish compared to those of their wildtype siblings under hypoxia. |