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. 2006 Jul 15;20(14):1885–1898. doi: 10.1101/gad.1424106

Figure 3.

Figure 3.

The N-terminal domain of GW182 interacts with the PIWI domain of AGO1. (A) HA-GW182 or the indicated protein fragments were transiently expressed in S2 cells. Cell lysates were immunoprecipitated using anti-HA antibodies. HA-MBP served as a negative control. Inputs and immunoprecipitates were analyzed by Western blot using anti-HA or anti-AGO1 antibodies. (B–G) Epitope-tagged GFP-AGO1 was expressed in S2 cells. In C–G, the effect of cotransfecting HA-tagged versions of GW182 or the indicated protein fragments on the localization of AGO1 was examined. The merged images show the GFP signal in green, the HA signal in red, and DNA in blue. Bar, 5 μm. (H) GST pull-down assays were performed with [35S]methionine-labeled full-length AGO1, AGO2, or the indicated AGO1 protein fragments, and recombinant GST or GST-GW182 (fragment 1–592). Samples were analyzed by SDS-PAGE followed by fluorography.