FIGURE 4.
Mutations in AGO1 that uncouple miRNA and GW182 binding. (A) Lysates from S2 cells expressing HA-tagged versions of MBP, wild-type AGO1, or AGO1 mutants were immunoprecipitated using a monoclonal anti-HA antibody. Inputs (1.5%) and immunoprecipitates (30%) were analyzed by Western blotting using a polyclonal anti-HA antibody. Endogenous GW182 was detected using anti-GW182 antibodies. The association between HA-AGO1 and endogenous bantam was analyzed by Northern blotting. tRNAAla served as a loading control. (Asterisk) IgG cross-reactivity.