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Fig. 1
Appl Proteins, Their Localization, and Biochemical Properties Are Conserved in Zebrafish
(A) Schematic representation of human and zebrafish APPL proteins indicating functional domains and total length.
(B and C) Zebrafish embryos injected with 100 pg appl1-Venus mRNA at single-cell stage and with 50 pg CFP-Rab5C mRNA into single blastomeres at 16-cell stage, fixed, and imaged at 50% epiboly stage. Single confocal sections are shown. Draq5 and phalloidin staining highlight nuclei and cell margins in blue.
(B) A cell coexpressing fluorescently labeled Appl1 and Rab5C proteins. Many labeled structures are shared between the two proteins (yellow, in merged panel, few highlighted by arrows). White arrowheads, structures that carry only Appl1-Venus; empty arrowheads, structures labeled by Rab5C only. The scale bar represents 10 μm.
(C) Distribution of ubiquitously expressed Appl1-Venus is altered in CFP-Rab5C coexpressing cells (outlined by white line): bigger and brighter structures are observed. Asterisk indicates cell represented in (B). The scale bar represents 20 μm.
(D) GST pull-down assays. Recombinant GST-Rab5C protein, loaded with either GTPγS (+GTP) or GDP (+GDP) nucleotides. In vitro translated Appl1 and Appl2 proteins bind specifically to Rab5C+GTP.
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