FIGURE 1.

Toll mRNA interacts with Wispy. Interactions of Toll mRNA with various cytoplasmic polyadenylation factors was assessed by RIP from Drosophila 90 min embryo extracts. (A) Orb2-GFP was immunoprecipitated with GFP-binder beads from Orb2-GFP, or OrR (wt) embryos used as negative control. Symplekin and Wispy were immunoprecipitated from OrR embryos, carrying nonspecific IgG as negative control (IgG). Fifteen percent (Symplekin, Wispy) or 50% (Orb2-GFP) of the pellet was loaded to visualize protein; i, 3%–10% input extract. (B) The level of Toll mRNA in the pellet was measured by RT-qPCR, normalized to the negative control, and represented as enrichment over immunoprecipitation of the nonpolyadenylation substrate sop (dashed line). The plot represents the average of at least three independent experiments. Error bars were calculated as standard deviation, and statistical significance analyzed by unpaired Student's t-test (*) P < 0.05; (**) P < 0.01; (***) P < 0.001. See Supplemental Table S1 for raw data and normalization details.