ZFIN Image: Vettori et al., 2017, Fig. S3

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Figure Caption

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.

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Acknowledgments

This image is the copyrighted work of the attributed author or publisher, and ZFIN has permission only to display this image to its users. Additional permissions should be obtained from the applicable author or publisher of the image. Full text @ Proc. Natl. Acad. Sci. USA