FIGURE

Fig. 5

ID

ZDB-FIG-080508-6

Publication

Schenck et al., 2008 - The endosomal protein Appl1 mediates Akt substrate specificity and cell survival in vertebrate development

Other Figures

All Figure Page

Back to All Figure Page

Fig. 5

Appl1 Regulates Akt Activity, Akt Substrate Specificity, and Akt-Dependent Survival in Zebrafish Development
(A) Western blot analyses of Akt and MAPK level and activity in WT, Appl1 knockdown (MO: 8 ng MO1A), Appl1 overexpressing (mRNA: 75 pg appl mRNA), and rescued embryos (MO + mRNA: 8 ng MO1A + 75 pg appl mRNA).
(B) Forebrain apoptosis assay: Quantitative evaluation of apoptosis in Appl1 knockdown embryos rescued by expression of Akt (Akt rescue: 8 ng MO1A + 100 pg Akt2 mRNA). Embryos were assigned to one of the four indicated cell death categories. Evaluation of Appl1 knockdown and appl1 mRNA-rescued embryos (Figure 4G) are shown for comparison. Total amount of embryos scored: n ^{Akt rescue} = 396. Error bars indicate standard deviation (SD) amongst five experiments.
(C) Western blot analyses on the same fish extracts analyzed in panel (A) monitoring level and activity of Gsk-3β and Tsc2.
(D) Western blot analyses of MOCK and LY294002-treated fish. (Ser9)-Gsk-3β and (Thr1462) phosphorylation critically depends on Akt activity.
(A–D) Experiments have been carried out on 54 hr zebrafish embryos.

Expression Data

Gene:	appl1
Antibodies:	Ab1-appl1 Ab1-gsk3b Ab1-tsc2 Ab1-tubg1 Ab2-akt Ab2-gsk3b Ab2-mapk Ab2-tsc2 Ab3-akt Ab5-mapk
Fish:	WT WT + MO1-appl1
Condition:	chemical treatment: LY294002
Knockdown Reagent:	MO1-appl1
Anatomical Term:	whole organism
Stage:	Long-pec

Expression Detail

Antibody Labeling

Phenotype Data

Fish:	WT + MO1-appl1
Knockdown Reagent:	MO1-appl1
Observed In:	forebrain
Stage:	Long-pec

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.

Reprinted from Cell, 133(3), Schenck, A., Goto-Silva, L., Collinet, C., Rhinn, M., Giner, A., Habermann, B., Brand, M., and Zerial, M., The endosomal protein Appl1 mediates Akt substrate specificity and cell survival in vertebrate development, 486-497, Copyright (2008) with permission from Elsevier. Full text @ Cell