hStaufen1 complexes are associated to the RISC proteins.
(A) Cultures of HEK293T cells were transfected with pChStaufen1-TAP (hStau-TAP) or pC-TAP (TAP) and either Ago1-HA, Ago2-HA, Ago3-HA or GFP-HA and soluble extracts were used for TAP purification. The purified complexes were analysed using antibodies specific for HA or hStau1. Total extract (Input), not bound to IgG (NBIgG), not bound to calmodulin (NBCa) and eluted with EGTA (Eluted) are shown. (B) Cultures of HEK293T cells were transfected with pChStaufen1-TAP or pC-TAP and the purified complexes were analysed with antibodies specific for hStau1 or Ago2. Total extract (Input), not bound to IgG (NBIgG) and eluted by digestion with TEV (Eluted) are shown. The mobilities of the Ago or GFP proteins or hStaufen-CBD are indicated to the right. Stars mark unspecific cross-reaction bands detected with the anti-hStau1 antibody and diamonds indicate endogenous hStau1 protein.