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. 2006 Jan 19;25(3):522–532. doi: 10.1038/sj.emboj.7600942

Figure 5.

Figure 5

PACT and TRBP coexist in the ∼500 kDa RISC. Total extract was prepared from HEK293T cells transiently transfected with FLAG-PACT, V5-Dicer, and V5-hAgo2 expression vectors and fractionated through a Sephacryl-S300 HR column. Each fraction was subject to immunoprecipitation with anti-FLAG antibody. The immunoprecipitate was used for (A) Western blot analysis using anti-TRBP or anti-FLAG antibody, or (B) in vitro RNAi assay using miR-30a-5p target RNA. The fraction number and the protein molecular mass standard (Sigma) are indicated at the top of the figures.