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Fig. 6. Eph^x652 mutants exhibit defects in dorsal lobe formation throughout MB development. (A,B) Adult MBs from wild-type (A) and Eph^x652 (B) individuals carrying UAS-mCD8:GFP driven by elav-Gal4 double-labeled with anti-GFP (green) and anti-Fas2 (red). Eph^x652 mutant has missing dorsal projections of both '/ß' (green) and /ß (yellow) MB neurons. (C,D) Pupal MBs from wild type (C) and Eph^x652 (D) stained with anti-Fas2 showing severely reduced dorsal lobe projections (arrow) in the Eph^x652 mutant. (E,F) Third instar MB from wild-type (E) and Eph^x652 (F) individuals carrying 201Y-Gal4 driving UAS-mCD8:GFP (green), which labels cell bodies of neurons (arrowhead in E) and their dorsal and medial axonal projections. Neurons show reduced dorsal lobe projections (arrow) in the Eph^x652 mutant (F). Reduction of dorsal lobe projections is accompanied by an increase in medial lobe projections (arrowhead in F). (G,H) Late stage embryos stained with anti-Fas2. In wild type (G), the developing MB neurons and their dorsal and medial projections are labeled (D, dorsal lobe; M, medial lobe; Ped, penduncle). (H) Eph^x652 mutant showing loss of dorsal lobe projections (asterisk). (I) MB of an Eph^x652/onecut^lx122 adult stained with anti-Fas2. Eph^x652/onecut^lx122 MBs are indistinguishable from wild type, showing no dorsal lobe defects. (J) MB of an Ephrin^KG09118 adult stained with anti-Fas2, showing defects in dorsal branch formation similar to Eph^x652. (K) Anti-Fas2 staining of an adult MB overexpressing Eph using OK107-Gal4 driving a UAS-Eph transgene. Similar to Eph^x652 loss-of-function mutants, dorsal lobes are absent, and medial ß projections are thicker (arrows), often appearing fused at the midline (arrowhead). (L) MB from an Eph^x652 adult stained with anti-Fas2 showing loss of dorsal lobes and thicker ß medial lobes (arrow), which are fused at the midline (arrowhead). Genotypes: (A) elav-Gal4 (X)/+ or Y;UAS-mCD8GFP/+; Eph^x652/CiD; (B) elav-Gal4 (X)/+ or Y;UAS-mCD8GFP/+;Eph^x652/Eph^x652; (C,G) Eph^x652/CiD; (D,H,K) Eph^x652/Eph^x652; (E) 201Y-Gal4/UAS-mCD8GFP;Eph^x652/CiD; (F) 201Y-Gal4/UAS-mCD8GFP;Eph^x652/Eph^x652; (I) Eph^x652/onecut^lx122; (J) Ephrin^KG09118/Ephrin^KG09118; (K) OK107-Gal4/+;UAS-Eph/+.

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