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. 2009 Feb 13;5(2):e1000374. doi: 10.1371/journal.pgen.1000374

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(A–G) 2 µm optical sections of larval brain hemispheres from late 3^rd instar larvae, displayed at the same scale. 20 µm scale bars. Frontal sections, midway through brains. Anterior up; midline to left. Glial cell nuclei are labeled with Repo (red); glial cell bodies and membranes are labeled with CD8GFP (green) driven by repo-Gal4; HRP (blue) reveals neuropil at high intensity and neuronal cell bodies at low intensity. repo>dMyc brains (C) contain many glia with enlarged nuclei (arrows). repo>dEGFR^λ;dMyc (D) brains have excess glia (red) throughout the brain, and these glia have very compact cell bodies (green) relative to wild-type glia (B). repo>dEGFR^λ;dCyclinD;dCdk4 (E) and repo>dEGFR^λ;Rbf1^dsRNA (F) brains display increased numbers of glia (red) relative to wild-type (B), but show a less severe glial phenotype and a more normal neuronal compartment (blue) than either repo>dEGFR^λ;dp110^CAAX (A) or repo>dEGFR^λ;dMyc (D). The combination of repo>Rbf1^dsRNA;dEGFR^λ;dp110^CAAX (G) enhances neoplasia, leading to a dramatic increase in aberrant glia. (H,I) dCyclinE expression (red) in repo>dEGFR^λ;dp110^CAAX glia (H) and in repo>dEGFR^λ;dp110^CAAX;Rbf1^dsRNA glia (I), alone and overlaid with the Repo glial marker (blue, right panels). dCyclinE is more strongly and more broadly expressed in neoplastic glia upon Rbf1 loss, as evidenced by the increased number and intensity of dCyclinE-expressing glial nuclei in (I) compared to (H). Anterior is up, midline to the left. 4.5 µm optical projections; 20 µm scale bars. (J) Pathway diagram of key effectors involved in glial neoplasia initiated by EGFR and PI3K, showing the pathway circuits driving cell cycle entry and progression, and protein translation. Arrows indicate pathway connections, although these connections are not necessarily direct. Genotypes: (A) UAS-dEGFR^λ UAS-dp110^CAAX/+; repo-Gal4 UAS-CD8GFP/+, (B) repo-Gal4 UAS-CD8GFP/+, (C) UAS-dMyc/repo-Gal4 UAS-CD8GFP, (D) UAS-dEGFR^λ/+; repo-Gal4 UAS-CD8GFP/UAS-dMyc, (E) UAS-dEGFR^λ/+; UAS-dCyclinD UAS-dCdk4/+; repo-Gal4 UAS-CD8GFP, (F) UAS-dEGFR^λ/+; repo-Gal4 UAS-CD8GFP/UAS-Rbf1^dsRNA, (G) UAS-dEGFR^λ UAS-dp110^CAAX/+; repo-Gal4 UAS-CD8GFP/UAS-Rbf1^dsRNA, (H) UAS-dEGFR^λ UAS-dp110^CAAX/+; repo-Gal4 UAS-CD8GFP/+, (I) UAS-dEGFR^λ UAS-dp110^CAAX/+; repo-Gal4 UAS-CD8GFP/UAS-Rbf1^dsRNA.