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Files in this Data Supplement:
Fig. S1. Ectopic expression of l(1)sc resulted in the disruption of lamina. Late L3 stage of control (A) and l(1)sc ectopically expressing (B) flies. Ectopic expression of l(1)sc was driven byNP6099 Gal4. Expression of Dac (magenta) and ap-lacZ (blue) are shown.
Fig. S2. Medullar development does not require retinal axon input in larval stage. (A,B) Lateral view (A) and horizontal section (B) of eya1 are shown. Expression of L(1)sc and Dpn was normal in eya1 mutant optic lobe but lamina was not formed. Expression of L(1)sc (green), Dpn (megenta) and Dac (white) are shown. Arrowhead in B indicates NE cell expressing L(1)sc. (C,D) Optic lobes of wild type (C) and eya1 (D) at 24-48 hours after puparium formation (APF) are shown. Excessive cell death and poorly organized smaller medulla neuropile were observed in eya1 animals. Cells undergoing apoptosis are visualized with cleaved caspase 3 antibody (green). Neuronal cell bodies, axons and neuropils are shown by anti-HRP antibody staining (magenta). La, lamina; Me, medulla; Lo, lobula.
Fig. S3. Expression pattern of upd-Gal4 and upd mRNA. (A) upd-Gal4 is expressed in lamina neurons in the late L3 stage. upd-Gal4 (green) is visualized by crossing with UAS-GFP.nls. Lamina neurons are visualized by immunostaining of Dac (magenta). (A′) Only upd-Gal4 expression is shown. (B-D) upd mRNA is specifically expressed in lateral NE cells. upd expression of early (B), mid (C) and late L3 stage (D) is shown (yellow arrowheads). (E) upd mRNA is specifically expressed in lamina furrow in the late L3 stage (horizontal section). Expression of upd mRNA (magenta) and Discs large (Dlg, blue) are shown. White arrowhead indicates lamina furrow.
Fig. S4. Late L3 optic lobe of Stat92E mutant. In Stat92E85C9/Stat92EF mutant, NBs disappeared, medulla neurons were fewer, and lamina was disrupted. Dac (green), Dpn (magenta), and Elav (blue) are shown.
Fig. S5. Cell growth does not alter progression of proneural wave and timing of medulla differentiation. (A) M(3)w124/+ cells are marked by GFP (blue in A) and +/+ cells by absence of GFP (circled by yellow line in A′). Expression of L(1)sc (green) and Dpn (magenta) was not altered. (B) M(3)w124/+ cells are marked by GFP (green in B) and +/+ cells by absence of GFP (circled by yellow line in B′). Expression of Dpn (magenta) and Arm (blue) was not altered.
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