Ago2 colocalizes with preannealed miRNA–mRNA duplexes in P-bodies. (A) Ago2 colocalizes with preannealed CXCR4–FL6X duplexes. HeLa cells were transiently transfected with HA-Ago2, and 24 h later, FL6X preannealed to Alexa568-labeled CXCR4 was transfected. After 16 h, immunostaining with Alexa488-labeled anti-HA antibody and DAPI was performed. (B–E) Ago2- and CXCR4-containing loci are P-bodies. HeLa cells were transfected with HA-Ago2 and immunostained 24 h later with antibodies against endogenous (B) GW182, (C) XRN1, (D) phospho-RPS6 (P-RPS6), and (E) RPL7a, followed by detection with Alexa647-labeled secondary antibodies. Relative fluorescence along the indicated paths was quantified using ImageJ and graphed as a function of distance.
