ZFIN ID: ZDB-FIG-171016-34
Vettori et al., 2017 - Glucocorticoids promote Von Hippel Lindau degradation and Hif-1α stabilization. Proc. Natl. Acad. Sci. USA   114(37):9948-9953 Full text @ Proc. Natl. Acad. Sci. USA
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Fish:
Condition:
Knockdown Reagent:
Anatomical Terms:
Stage: Protruding-mouth
PHENOTYPE:
Fish:
Condition:
Knockdown Reagent:
Observed In:
Stage: Protruding-mouth

Fig. S3

DEX induces Hif-1 pathway activation and expression of glycolytic regulators and is GR-dependent. (A) Histograms showing the average values of the integrated density of fluorescence in 72-hpf Tg(4xhre-tata:eGFP)ia21 embryos treated with different doses of DEX. DEX incubation induces a dose-dependent activation of the Hif-1 pathway as shown by the increase of the 4xhre-tata transgene expression. A.U., arbitrary units. (B) In situ hybridization for glycolytic regulators ldha, glut1, and pfkfb3 showing increased expression after DEX treatment. (Magnification: 17.5×.) (C) Fold changes in gene expression of fkbp5 in 72-hpf embryos injected with gr-MO and CTRL-MO compared with nontreated/noninjected control (set at 1). Real-time PCR analysis revealed that the expression of fkbp5 was significantly reduced in gr-MO–injected embryos, confirming the ability of gr-MO to impair the expression of the glucocorticoid receptor. (D, Left) Images of 72-hpf Tg(4xhre-tata:eGFP)ia21 in wild-type and GR CRISPR mutant background treated with 10 μM DEX in DMSO for 24 h or with vehicle control. (Magnification: 10×.) (Right) Quantification of GFP reporter fluorescence, showing that the HRE reporter failed to respond in the GR mutant background. Generation of this GR mutant is described in SI Text.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
EGFP ia21Tg control Protruding-mouth whole organism IFL
ia21Tg chemical treatment: dexamethasone Protruding-mouth whole organism IFL
ia21Tg ; nr3c1ia30/ia30 control Protruding-mouth whole organism IFL
ia21Tg ; nr3c1ia30/ia30 chemical treatment: dexamethasone Protruding-mouth whole organism IFL
fkbp5 ia21Tg control Protruding-mouth whole organism RTPCR
ia21Tg + MO4-nr3c1 control Protruding-mouth whole organism RTPCR
ldha ia21Tg control Protruding-mouth anatomical structure ISH
ia21Tg chemical treatment: dexamethasone Protruding-mouth anatomical structure ISH
pfkfb3 ia21Tg control Protruding-mouth anatomical structure ISH
ia21Tg chemical treatment: dexamethasone Protruding-mouth anatomical structure ISH
slc2a1a ia21Tg control Protruding-mouth anatomical structure ISH
ia21Tg chemical treatment: dexamethasone Protruding-mouth anatomical structure ISH
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
ia21Tg chemical treatment: dexamethasone Protruding-mouth anatomical structure pfkfb3 expression increased amount, abnormal
Protruding-mouth anatomical structure ldha expression increased amount, abnormal
Protruding-mouth anatomical structure slc2a1a expression increased amount, abnormal
Protruding-mouth whole organism EGFP expression increased amount, abnormal
ia21Tg + MO4-nr3c1 control Protruding-mouth whole organism fkbp5 expression decreased amount, abnormal
ia21Tg ; nr3c1ia30/ia30 chemical treatment: dexamethasone Protruding-mouth whole organism EGFP expression amount, ameliorated
Acknowledgments:
ZFIN wishes to thank the journal Proc. Natl. Acad. Sci. USA 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