Ligand-Independent Traffic of Notch Buffers Activated Armadillo in Drosophila (original) (raw)
Figure 4
Localization of CeN, CN, and Ce in the epithelial cells of third instar wing discs.
(A–A3) Localization of CeN at and above the adherens junctions (insets in A and A1 show the adherens junctions as labelled by DCad staining in the same image) as well as in more basal dots, which represent vesicles (A2–A3). The localization is revealed by the fluorescence of the eGFP. Notice that there is a pool of Notch apical to the adherens junctions. (B–B2) Localization of Ce in all membranes at all levels of the cell (B, apical' B1, subapical; B2, basal). This molecule contains the eGFP fused to the transmembrane and extracellular domains of CD8 and indicates that the localization of CeN is determined by the sequences of the Notch protein that are added to Ce. (C–E) Confocal optical z-sections through the wing pouch of a disc expressing CeN, showing the large vesicles of stain (C); CN, a chimera like CeN but without the eGFP that can only be visualized with an antibody against the intracellular domain of Notch, NICD, (D); and Ce, highlighting the overall and nonspecific distribution of eGFP to all membranes of the cell (E). Notice that in (D) it is not easy to distinguish the endogenous Notch from CN other than by the amount. The arrows in (C) and (E) point to the approximate levels of the pictures (A–A3) and (B–B2). The expression of the different proteins is directed by _dpp_-Gal4 in all cases. Scale bar, 10 µm. (F) Schematic of the structure of the Notch receptor and related chimeras used in this work.