Fig. 5. The Min protein localises to the apical membrane at the time of cuticulin envelope assembly. (A,B) Confocal image of a transverse section taken from wild-type pupal legs stained for Min protein. (A) At 6 hours APF, Min protein accumulates in the cytoplasm (n, cell nuclei), whereas the staining accumulates in the cells apical surface 1 hour later (B). The position of the basal and apical surfaces is indicated. (C,D) Confocal images corresponding to an apical plane of a slightly tilted pupal wild-type wings. (C) At 32 hours APF, Min (green) accumulates in the cytoplasm, both apically (arrowheads, upper part of the picture) or in a more basolateral focal plane (arrows, bottom of the picture). (D) At 34 hours APF, a uniform layer stained with the anti-Min antibody covers the whole wing and the epidermal hairs (arrowheads). Anti E-cadherin antibodies (red) mark the position of apical junctions. (E-F) Transverse section of similar wings to those shown in B. Strong expression of -catenin—GFP (red) marks both apical junctions and the basal part of the cells (white line). (E) At 32 hours APF, Min protein accumulates in the apical side of the cell and in the cytoplasm (green). Nuclei position is marked by the TOPRO dye (blue). The dorsal and ventral cell layers composing the wing are evident; the basal membrane is now in the middle of the wing (white line). (F) At 34 hours APF, Min protein is detected only in the apical side of the cells. (H,I) Confocal view of the wing margin row of stout bristles stained with anti-Min. (H) By 32 hours APF, most of the staining accumulates in the cell body of the trichogen cells (tr), whereas the bristle shafts are not stained (arrowheads). (I) 2 hours later the staining has disappeared from the cytoplasm of the trichogen cells (tr) and accumulates in the periphery of the growing shaft (arrowheads). The tormogen cells (to) of each bristle still accumulate Min in their cytoplasm at this stage.

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