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Figure 1


Fig. 1. (A) Expression and post-translational modifications of [S239] and [C93A]Wingless. Drosophila S2 cells were transfected with plasmids expressing either the wild-type or the [C93A]Wingless protein and processed for immunoblotting (left lanes). Right-hand-side lane shows [S239]Wingless obtained from transgenic imaginal discs. Distinct bands representing different states of glycosylation can be detected in all three cases. (B) [C93A]Wingless has no significant signalling activity, and that of [S239]Wingless is strongly reduced. Drosophila S2 cells were transfected with the topflash reporter along with plasmids expressing Fz2 and Renilla luciferase. The cells were also transfected with equal amounts of plasmids expressing [S239]Wingless, [C93A]Wingless or wild-type Wingless. Luciferase activity was measured and normalised against Renilla activity for the two groups of cells. (C) Diagram of a wing imaginal disc showing the domain of Wingless expression in green and of ap-Gal4 in red. (D-F) Expression of wild-type Wingless (D), [S239]Wingless (E) and [C93A]Wingless (F) under the control of ap-Gal4. Unlike the wild-type protein, the mutant proteins cannot be detected in non-expressing cells (no punctate staining can be detected). Scale bar: 10 µm. (G-N) Extracellular staining versus total staining of various Wingless forms expressed under the control of ap-Gal4. In all panels, total staining is shown in red and extracellular staining in green. Scale bar: 50 µm. (G,K) Expression of wild-type Wingless. Note the presence of extracellular signal at the surface of expressing cells, in the dorsal compartment (square brackets). The pouch is enlarged because of excess activation of Wingless signalling. (H,L) Expression of [S239]Wingless. Strong extracellular staining can be detected in the expression domain. (I,M) Expression of [C93A]Wingless. No extracellular signal is detected. (J,N) Expression of NRT-[C93A]Wingless. Extracellular localisation is restored by the presence of a heterologous transmembrane domain.





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