Figure 3.

miRNA target association with eIF4E is rescued in cells depleted of decapping factors. (A–G) Control S2 cells (treated with GFP dsRNA) or cells depleted of DCP1 and EDC4 were transfected with a mixture of three plasmids as described in Figure 1. (F, G) The transfection mixtures additionally contained plasmids for the expression of the indicated HA-tagged proteins. (A) Firefly luciferase activity was normalized to that of the Renilla luciferase. For each condition (control and knockdown cells), the normalized values of F-Luc activity were set at 100 in the absence of miR-9b (white bars). (B) Northern blot of representative RNA samples analysed as described in Figure 1. The positions of the polyadenylated (A_n) and deadenylated (A₀) F-Luc-Nerfin-1 mRNA are indicated. (C–E) Coimmunoprecipitation of the F-Luc-Nerfin-1 reporter with the endogenous indicated proteins in the absence (white bars) or presence of miR-9b (blue bars). (F, G) Coimmunoprecipitation of the F-Luc-Nerfin-1 reporter with HA-tagged proteins in the absence (white bars) or presence of miR-9b, analysed as described in Figure 1.

Source data for this figure is available on the online supplementary information page.

Source Data for Figure 3 ^{(980.8KB, pdf)}