FIGURE

Fig. 4

ID
ZDB-FIG-101111-44
Publication
Prykhozhij, 2010 - In the Absence of Sonic Hedgehog, p53 Induces Apoptosis and Inhibits Retinal Cell Proliferation, Cell-Cycle Exit and Differentiation in Zebrafish
Other Figures
All Figure Page
Back to All Figure Page
Fig. 4

(A) p53 reporter PG13p21::nlsEGFP-polyA was constructed using Tol2kit as described in Materials and methods. It consists of the p21 promoter fused with p53-binding enhancer sequence PG13, nlsEGFP coding sequence and a polyadenylation signal (polyA). (B) Wild-type embryos carrying the p53 reporter transgene were incubated for 6 hours at 24 hpf with either 50 μM Roscovitine or 0.4% DMSO diluted in E3 medium. DMSO-treated embryos did not show any significant expression of the reporter (right), whereas roscovitine-treated ones expressed p53 reporter at a high level (left). All embryos showed green-heart transgenesis marker. The experiment was repeated three times and 20 embryos were used for each treatment. (C) p53 morpholino but not 4-mismatch control morpholino injection suppressed expression of PG13p21::nlsEGFP p53 reporter in shh-/- mutant embryos at 24 hpf. Both injections had no effect on p53 reporter expression in the wild-type embryos. (D) Anti-active Caspase3 staining of cells in the shh-/- mutant retina at 56 hpf expressing p53 reporter transgene PG13p21::nlsEGFP. Most cells positive for active Caspase3 also expressed p53 reporter transgene. Single channel, overlay and co-localisation analysis images are shown. (E) Quantification of co-localisation of active Caspase3 staining and nlsEGFP p53 reporter signal. Co-localised pixels made up 20,9 +/- 3,8% of total active Caspase3 pixels and 11,7 +/ 4,5% of total nlsEGFP pixels. Quantifications were performed on 20 retinal section images from 10 embryos. Asterisks (***) indicate a significant statistical difference between the proportions (t-test, P-value<0,001).

Expression Data

Expression Detail
Antibody Labeling
Phenotype Data

Phenotype Detail
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 @ PLoS One