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

Figure 6.

Figure 6

PACT is required for siRNA-induced RNAi. (A) HeLa cells were transiently transfected with siRNA targeting Dicer, PACT, or TRBP along with siRNA against firefly luciferase, firefly luciferase-expression plasmid, and renilla luciferase-expression plasmid. The firefly luciferase activity was normalized against renilla luciferase activity. This experiment was performed in triplicate. Error bars indicate standard deviations. (B) Western blot analysis. siRNA targeting GFP, Dicer, hAgo2, TRBP, or PACT was transfected into HeLa cells. Following transfection (48 h), total extract was prepared in lysis buffer (10 mM Tris, pH 7.4, 1 mM EDTA, 500 mM NaCl, 0.5% Triton-X100). In all, 120 μg of total extract was loaded on 10% SDS–PAGE. Endogenous Dicer, TRBP, or PACT was visualized using anti-Dicer, anti-TRBP, or anti-PACT antibody.