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. 2005 Apr;25(8):3027–3039. doi: 10.1128/MCB.25.8.3027-3039.2005

FIG. 7.

FIG. 7.

dDP and dE2F2 levels are interdependent in mutant animals and in cells. (A) Cells were treated with nonspecific (NS), dE2F1 (E1), dDP, and dE2F2 (E2) dsRNAs, and cell extracts were analyzed on Western blots using antibodies specific for dE2F1, dE2F2, and dDP. Note that depletion of dDP affects the levels of both dE2F1 and dE2F2, and conversely, dDP is reduced following depletion of dE2F1 and dE2F2. Anti-tubulin mouse monoclonal antibody was used as a loading control. (B) Western blot analysis of larval extracts from Canton S (wild-type), de2f276Q1/de2f2G5.1 (de2f2−), de2f276Q1/de2f2G5.1, de2f191/de2f1rm729 (de2f2−; de2f1−), and dDPa2/Df(2R)vg-B (dDP−) animals. (C) A wild-type level of dDP can be restored in de2f2 mutants by the reexpression of dE2F2. A heat shock-inducible de2f2 transgene, hs-de2f2, was introduced into de2f2 mutant animals. The level of dDP in larvae was monitored by Western blot analysis before heat shock (no HS) and at 30 min (30′ AHS) and 2 h (2 h AHS) following a 1-h heat shock (de2f2− hs-de2f2). The rescue was not due to the heat shock treatment, because the level of dDP was unchanged in de2f2 mutants that lack an hs-de2f2 transgene (de2f2−). (D) Northern blot analysis of larvae of the same genotypes used in panel B. 32P-labeled RNA probes for de2f1, de2f2, and dDP reveal that there is no decrease in the steady-state level of dDP mRNA in the de2f1 de2f2 double mutant or of de2f1 and de2f2 mRNAs in the dDP mutant.