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. 2010 Jun 14;189(6):981–996. doi: 10.1083/jcb.200910006

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© 2010 Buttitta et al.

This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 3.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/3.0/).

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Figure 5. — APC/C activity limits E2F-induced bypass of exit. (A) Several APC/C components (APC1, -4, -5, -6, -7, -8, and -10) and regulators (cdc20, cdc20-like, and Cks30A) are transcriptionally increased (1.5–40-fold) when E2F activity is high or de-repressed in Dp mutant wings. (B–I) GFP-marked clones expressing the indicated cell cycle regulators were generated using hs-FLP tub>Gal4/UAS, tub-Gal80^TS and examined for PH3 (B–I) or MPM2 (F). Nuclei were stained with Hoechst (blue; G–I). PH3 is seen in clones expressing the APC/C inhibitor Rca1 at 24–36 h APF in the eye and wing (wing shown; B) but not the late wing and was only observed once in the eye at 40–44 h APF (C, arrow). When E2F1–DP and Rca1 are coexpressed, bypass of exit is observed as late as 60 h APF (D) without disrupting terminal differentiation of ommatidia or bristles (E). Yellow outline indicates clone boundary. Coexpression of E2F1–DP with Fzy RNAi or Fzr RNAi also led to ectopic mitoses in eyes (F and H) and wings (G and I) at 48 h APF, as shown by PH3 staining (F–I) and MPM2 cytoplasmic mitotic staining (F; white in F′ inset; arrowhead indicates cytoplasmic MPM2 staining). G1-S progression was also observed in clones expressing E2F1–DP with Fzy RNAi in eyes at 48 h APF, as indicated by MPM2 nuclear foci (F; white in F′ inset; arrows point to foci). Sparse clones overexpressing E2F + RNAis for the three APC/C activators were induced at 0 h white prepupae, and cells per clone for at least 50 clones/genotype were quantified at 54–56 h APF. The distributions of clonal cell counts and mean cells/clone are indicated in J. (J, right) Examples of large clones and overlying wing cuticle and hairs at 56 h APF. DIC, differential interference contrast. Bars, 50 µm.