ZFIN ID: ZDB-FIG-171016-30
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:
Conditions:
Knockdown Reagent:
Anatomical Terms:
Stage Range: Protruding-mouth to Day 5
PHENOTYPE:
Fish:
Conditions:
Knockdown Reagent:
Observed In:
Stage Range: Protruding-mouth to Day 5

Fig. 1

Identification of glucocorticoids as HIF activators. (A) Retest of hits from the Spectrum screen for activators of the HIF response in phd3:eGFP zebrafish, where compounds showing an average GFP level of >44 are shown. Averaged maximum pixel intensity and SEM are shown for between five and nine embryos. Dark gray: controls; black: GC-like compounds; light gray: non-GC hits. AMC, amcinonide; BME, betamethasone; CBP, clobetasol propionate; CPO, ciclopirox olamine; DEX, dexamethasone; DFL, 8,2-dimethoxyflavone; FLC, fluocinonide; HAL, halcinonide; HCB, hydrocortisone butyrate; ICA, icariin; NIN, 7-nitroindazole; PTE, ptaeroxylin; TAA, triamcinolone acetonide; TAD, triamcinolone diacetate. (B) BME activates phd3:eGFP in an RU-486–dependent manner; representative images show the activation specific to the liver in BME-treated embryos and reduced GFP expression following BME+RU-486 cotreatment. (Magnification: 16×.) (C) Quantitation of phd3:eGFP response. DMSO was added to 1% vol/vol in all experiments except Control (C), DMOG to 60 μM, and RU-486 to 5 μM as indicated (n ≥ 9). (D) BME induction of hypoxia-regulated genes and GC target genes, where values above bars indicate average fold change (FC) and triplicate samples of 20 embryos were used. For the hypoxia-induced genes, FC induction in vhl mutants vs. wild type is given as a comparison, where FC is calculated using ΔΔCT ± SEM. (E) HIF1β-MO abrogates BME activation of phd3:eGFP. Maximum pixel intensity in embryos as grouped by injection showing mean ± SEM (n = 14). Significance was calculated using Kruskal–Wallis one-way ANOVA followed by Dunn’s multiple comparison test for treatments against DMSO (*P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001).

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
EGFP sh144Tg/sh144Tg control Protruding-mouth whole organism IFL
Day 5 liver IFL
Day 5 whole organism IFL
sh144Tg/sh144Tg chemical treatment: amcinonide Day 5 whole organism IFL
sh144Tg/sh144Tg chemical treatment: betamethasone Protruding-mouth whole organism IFL
Day 5 liver IFL
Day 5 whole organism IFL
sh144Tg/sh144Tg chemical treatment: betamethasone, chemical treatment: mifepristone Day 5 liver IFL
sh144Tg/sh144Tg chemical treatment: clobetasol propionate Day 5 whole organism IFL
sh144Tg/sh144Tg chemical treatment: ciclopirox olamine Day 5 whole organism IFL
sh144Tg/sh144Tg chemical treatment: dexamethasone Day 5 whole organism IFL
sh144Tg/sh144Tg chemical treatment: 8-methoxy-2-(2-methoxyphenyl)-1-benzopyran-4-one Day 5 whole organism IFL
sh144Tg/sh144Tg chemical treatment: dimethyloxalylglycine Day 5 whole organism IFL
sh144Tg/sh144Tg chemical treatment: dimethyloxalylglycine, chemical treatment: mifepristone Day 5 whole organism IFL
sh144Tg/sh144Tg chemical treatment: Fluocinonide Day 5 whole organism IFL
sh144Tg/sh144Tg chemical treatment: Halcinonide Day 5 whole organism IFL
sh144Tg/sh144Tg chemical treatment: cortisol 17-butyrate Day 5 whole organism IFL
sh144Tg/sh144Tg chemical treatment: icariin Day 5 whole organism IFL
sh144Tg/sh144Tg chemical treatment: 7-NITROINDAZOLE Day 5 whole organism IFL
sh144Tg/sh144Tg chemical treatment: 5-hydroxy-2,8-dimethyl-6,9-dihydropyrano[3,2-h][1]benzoxepin-4-one Day 5 whole organism IFL
sh144Tg/sh144Tg chemical treatment: mifepristone Day 5 whole organism IFL
sh144Tg/sh144Tg chemical treatment: triamcinolone acetonide Day 5 whole organism IFL
sh144Tg/sh144Tg chemical treatment: Triamcinolone diacetate Day 5 whole organism IFL
sh144Tg/sh144Tg + MO1-arnt control Protruding-mouth whole organism IFL
sh144Tg/sh144Tg + MO1-arnt chemical treatment: betamethasone Protruding-mouth whole organism IFL
fkbp5 sh144Tg/sh144Tg control Day 5 whole organism RTPCR
sh144Tg/sh144Tg chemical treatment: betamethasone Day 5 whole organism RTPCR
sh144Tg/sh144Tg ; vhlhu2117/hu2117 control Day 5 whole organism RTPCR
sh144Tg/sh144Tg ; vhlhu2117/hu2117 chemical treatment: betamethasone Day 5 whole organism RTPCR
hpxa sh144Tg/sh144Tg control Day 5 whole organism RTPCR
sh144Tg/sh144Tg chemical treatment: betamethasone Day 5 whole organism RTPCR
sh144Tg/sh144Tg ; vhlhu2117/hu2117 control Day 5 whole organism RTPCR
sh144Tg/sh144Tg ; vhlhu2117/hu2117 chemical treatment: betamethasone Day 5 whole organism RTPCR
p4ha1b sh144Tg/sh144Tg control Day 5 whole organism RTPCR
sh144Tg/sh144Tg chemical treatment: betamethasone Day 5 whole organism RTPCR
sh144Tg/sh144Tg ; vhlhu2117/hu2117 control Day 5 whole organism RTPCR
sh144Tg/sh144Tg ; vhlhu2117/hu2117 chemical treatment: betamethasone Day 5 whole organism RTPCR
pck1 sh144Tg/sh144Tg control Day 5 whole organism RTPCR
sh144Tg/sh144Tg chemical treatment: betamethasone Day 5 whole organism RTPCR
sh144Tg/sh144Tg ; vhlhu2117/hu2117 control Day 5 whole organism RTPCR
sh144Tg/sh144Tg ; vhlhu2117/hu2117 chemical treatment: betamethasone Day 5 whole organism RTPCR
pfkfb3 sh144Tg/sh144Tg control Day 5 whole organism RTPCR
sh144Tg/sh144Tg chemical treatment: betamethasone Day 5 whole organism RTPCR
sh144Tg/sh144Tg ; vhlhu2117/hu2117 control Day 5 whole organism RTPCR
sh144Tg/sh144Tg ; vhlhu2117/hu2117 chemical treatment: betamethasone Day 5 whole organism RTPCR
scarb1 sh144Tg/sh144Tg control Day 5 whole organism RTPCR
sh144Tg/sh144Tg chemical treatment: betamethasone Day 5 whole organism RTPCR
sh144Tg/sh144Tg ; vhlhu2117/hu2117 control Day 5 whole organism RTPCR
sh144Tg/sh144Tg ; vhlhu2117/hu2117 chemical treatment: betamethasone Day 5 whole organism RTPCR
tsc22d3 sh144Tg/sh144Tg control Day 5 whole organism RTPCR
sh144Tg/sh144Tg chemical treatment: betamethasone Day 5 whole organism RTPCR
sh144Tg/sh144Tg ; vhlhu2117/hu2117 control Day 5 whole organism RTPCR
sh144Tg/sh144Tg ; vhlhu2117/hu2117 chemical treatment: betamethasone Day 5 whole organism RTPCR
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
sh144Tg/sh144Tg chemical treatment: amcinonide Day 5 whole organism EGFP expression increased amount, abnormal
sh144Tg/sh144Tg chemical treatment: betamethasone Protruding-mouth whole organism EGFP expression increased amount, abnormal
Day 5 liver EGFP expression increased amount, abnormal
Day 5 whole organism fkbp5 expression increased amount, abnormal
Day 5 whole organism pck1 expression increased amount, abnormal
Day 5 whole organism EGFP expression increased amount, abnormal
Day 5 whole organism pfkfb3 expression increased amount, abnormal
Day 5 whole organism p4ha1b expression increased amount, abnormal
Day 5 whole organism scarb1 expression increased amount, abnormal
Day 5 whole organism hpxa expression increased amount, abnormal
Day 5 whole organism tsc22d3 expression increased amount, abnormal
sh144Tg/sh144Tg chemical treatment: betamethasone, chemical treatment: mifepristone Day 5 liver EGFP expression amount, ameliorated
sh144Tg/sh144Tg chemical treatment: clobetasol propionate Day 5 whole organism EGFP expression increased amount, abnormal
sh144Tg/sh144Tg chemical treatment: ciclopirox olamine Day 5 whole organism EGFP expression increased amount, abnormal
sh144Tg/sh144Tg chemical treatment: dexamethasone Day 5 whole organism EGFP expression increased amount, abnormal
sh144Tg/sh144Tg chemical treatment: 8-methoxy-2-(2-methoxyphenyl)-1-benzopyran-4-one Day 5 whole organism EGFP expression increased amount, abnormal
sh144Tg/sh144Tg chemical treatment: dimethyloxalylglycine Day 5 whole organism EGFP expression increased amount, abnormal
sh144Tg/sh144Tg chemical treatment: dimethyloxalylglycine, chemical treatment: mifepristone Day 5 whole organism EGFP expression increased amount, abnormal
sh144Tg/sh144Tg chemical treatment: Fluocinonide Day 5 whole organism EGFP expression increased amount, abnormal
sh144Tg/sh144Tg chemical treatment: Halcinonide Day 5 whole organism EGFP expression increased amount, abnormal
sh144Tg/sh144Tg chemical treatment: cortisol 17-butyrate Day 5 whole organism EGFP expression increased amount, abnormal
sh144Tg/sh144Tg chemical treatment: icariin Day 5 whole organism EGFP expression increased amount, abnormal
sh144Tg/sh144Tg chemical treatment: 7-NITROINDAZOLE Day 5 whole organism EGFP expression increased amount, abnormal
sh144Tg/sh144Tg chemical treatment: 5-hydroxy-2,8-dimethyl-6,9-dihydropyrano[3,2-h][1]benzoxepin-4-one Day 5 whole organism EGFP expression increased amount, abnormal
sh144Tg/sh144Tg chemical treatment: triamcinolone acetonide Day 5 whole organism EGFP expression increased amount, abnormal
sh144Tg/sh144Tg chemical treatment: Triamcinolone diacetate Day 5 whole organism EGFP expression increased amount, abnormal
sh144Tg/sh144Tg + MO1-arnt chemical treatment: betamethasone Protruding-mouth whole organism EGFP expression amount, ameliorated
sh144Tg/sh144Tg ; vhlhu2117/hu2117 chemical treatment: betamethasone Day 5 whole organism pfkfb3 expression increased amount, abnormal
Day 5 whole organism scarb1 expression increased amount, abnormal
Day 5 whole organism tsc22d3 expression increased amount, abnormal
Day 5 whole organism fkbp5 expression increased amount, abnormal
Day 5 whole organism hpxa expression increased amount, abnormal
Day 5 whole organism pck1 expression increased amount, abnormal
Day 5 whole organism p4ha1b expression increased amount, abnormal
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