Fig 1: Coimmunoprecipitations of candidate interacting factors. (A) Co-IP of AGO2 with RNAi factors from the nucleus. AGO2 was immunoprecipitated versus an IgG control and interacting factors were blotted for with antibodies by Western blot. (B) Co-IP of Flag-AGO2 with interacting factors from the nucleus identified by mass spectrometry. Flag-AGO2 was immunoprecipitated from Flag-AGO2 stable line cells versus a Flag IP from endogenous T47D cells. Interacting factors were blotted for with antibodies by Western blot. (C) Co-IP of AGO2 with potential interacting factors from the nucleus. AGO2 was immunoprecipitated versus an IgG control and interacting factors were blotted for with antibodies by Western blot. (D) Model of nuclear AGO2 activity. AGO2 is bound by TRNC6 proteins and AGO3 in the cytoplasm. AGO2 is loaded by DICER, TRBP, and other loading factors. Active RISC is then shuttled into the nucleus. Interactions with DICER and TRBP are more transient in the nucleus. Loaded AGO2 and TNRC6 proteins can then alter transcription or splicing within the nucleus, depending on the small RNA bound by AGO2.