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. Author manuscript; available in PMC: 2013 Nov 1.
Published in final edited form as: Dev Dyn. 2012 Sep 17;241(11):1695–1707. doi: 10.1002/dvdy.23857

FIGURE 6. Reducing the levels of ORC5 restores dE2F1 activity but this is independent of dE2F1 protein levels.

FIGURE 6

6A) Third instar larvae wing discs expressing a PCNA-GFP reporter construct were immunostained with antibodies against GFP (6Aa-d) or probed for PCNA expression by in situ hybridization (6Ba-d). Larvae expressing de2f1-dsRNA driven by ptc-GAL4 (Ab and Bb) exhibit a reduction in PCNA in the area of the ptc-GAL4 stripe (arrow) when compared to wild-type (Aa, Ba). The reduction in PCNA staining was suppressed when Orc5-dsRNA was expressed concomitantly (Ac, Bc). Discs expressing Orc5-dsRNA alone showed no effect on PCNA expression (Ad, Bd).

6C) Immunofluorescent staining of third instar wing discs using antibodies against dE2F1 (red) and GFP (green, to mark the ptc-GAL4 stripe, a’-d’) demonstrate that larvae which express de2f1-dsRNA show a reduction of dE2F1 protein levels (b), when compared to wild-type (a). Reducing the levels of ORC5 simultaneously did not interfere with the targeting of dE2F1 (c) and expression of Orc5-dsRNA alone did not change the overall levels of dE2F1 (d).