Mammalian GW182 contains multiple Argonaute-binding sites and functions in microRNA-mediated translational repression

Each Ago2-binding domain of GW182 binds to an overlapping region of Ago2. (A) HEK293F cells were cotransfected with the FLAG/SBP-GW182 445–526, FLAG-GW182 560–806, or 778–910 expression vector, with or without the FLAG–Ago2 expression vector. (B) HEK293F cells were cotransfected with the FLAG/SBP-GW182 560–806 and FLAG-GW182 778–910 expression vectors, with or without the FLAG-Ago2 expression vector. SBP pull-down assays and Western blotting were performed as described in Figure 1C, in the presence of RNaseA.