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Fig. 5. Overexpressed Dx increases the number of endocytic vesicles containing N and stabilizes the N in these vesicles. (A-L) The expression of N⁺-GV3 protein (green) in eye discs was detected by anti-GAL4. Dx was driven by the GMR promoter (purple in I-L). (A,E,I) Wild-type (A) and GMR-dx (E,I) eye discs before heat shock. No expression of N⁺-GV3 (green) was detected. I is a merged image. (B,C) In wild-type eye discs, N⁺-GV3 expression (green) was observed throughout the eye disc, except for the morphogenetic furrow, 30 minutes after heat shock. The N⁺-GV3 expression was primarily localized to the plasma membrane and within a small number of intracellular vesicles. C is a higher magnification of B. (D) In the wild-type eye disc, no N⁺-GV3 expression (green) was observed 12 hours after heat shock. (F,G,J,K) In GMR-dx eye discs, N⁺-GV3 expression (green) was observed throughout the eye disc, except for the morphogenetic furrow, 30 minutes after heat shock. In the region expressing Dx (purple), N⁺-GV3 expression was primarily localized to the intracellular vesicles. G and K are a higher magnification of F and J, respectively. J and K are merged images. (H,L) In the region expressing Dx (purple), N⁺-GV3 (green) was still detected. L is a merged image. (M-Q) In situ hybridization of wild-type (M,N,Q) and GMR-dx (O,P) eye discs using antisense (M-P) and sense (Q) GAL4 probes. N⁺-GV3 mRNA was detected using the antisense GAL4 probe. (M,O) In wild-type and GMR-dx eye discs, N⁺-GV3 mRNA was observed throughout the eye disc 30 minutes after heat shock. (N,P) In wild-type and GMR-dx eye discs, no N⁺-GV3 mRNA was detected 12 hours after heat shock. (Q) In situ hybridization using a sense-strand probe for GAL4. (R,S) Within the dx^–/dx^– clones (patches without green fluorescence), the number of vesicles containing N⁺-GV3 (purple) was reduced, compared with wild-type and dx^–/+ cells. (T) The number of vesicles containing N⁺-GV3 per cell was compared in seven independent measurements. The relative number of vesicles in the dx^–/dx^– cells are shown as percentages.

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