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Fig. 3. Echinoid subcellular localisation and colocalisation with Notch and Delta. Third instar larval discs examined by confocal microscopy. (A) Ed protein in wild-type wing disc. (B) sca-Gal4/UAS-ed-GFP. Detection of Ed-GFP (green) and Ed (red). (C) sca-Gal4/UAS-fng-DXDmut-Myc. Ed (red) and Golgi represented by Myc (green). Arrows indicate some Ed protein that is in the Golgi. Arrowheads indicate some of the more numerous Ed-only vesicles. (D) Immunohistochemical detection of Sca (green) and Ed (red), showing lack of colocalisation. (A-D) Single confocal sections. (E) Immunohistological detection of Ed (green), HRS (red) and N^ECD (blue). Confocal xzy projection. Arrows indicate vesicles containing all three proteins. (E') Green channel (Ed). (E'') Red channel (N^ECD). (E''') Blue channel (HRS). (F,G) Ed expression in confocal projections. (F) Wild type. (G) hook¹ mutant, showing fewer Ed-positive vesicles. (H,I) Immnuohistochemical detection of Dl (green) and Ed (red). Single confocal sections of wild-type discs. (H) Wing disc. Note that the majority of the Ed colocalises with Dl; some of the Ed- and Dl-positive vesicles are marked by arrows. Dl-only vesicles are marked by arrowheads. (I) Eye disc. Ed does not colocalise very strongly with Dl. (J) Immunohistochemical detection of Ed-GFP (green), N^ECD (red) and Dl (blue). Confocal xzy projection. Arrows indicate vesicles containing all three proteins. (J') Green channel (Ed-GFP). (J'') Red channel (N^ECD). (J''') Blue channel (Dl).

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