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Figure 2 | BMC Molecular Biology

Figure 2

From: Characterization of the TRBP domain required for Dicer interaction and function in RNA interference

Figure 2

The C4 domain is necessary and sufficient to mediate TRBP-Dicer interaction. A) TRBP1 or TRBP1ΔC4-Dicer interactions by immunoprecipitation. tarbp2-/- cells were transfected with none (lanes 1, 4, 7) or 15 μg of pCMV-Myc-TRBP1 (lanes 2, 5, 8), pCMV-Myc-TRBP1ΔC4 (lanes 3, 6, 9). IP was performed with 1 mg of protein and anti-Myc antibody (lanes 4–6) or anti-Dicer antibody (lanes 7–9). 100 μg of proteins from each lysate (input; lanes 1–3) and the immunoprecipitated complexes (lanes 4–9) were run on a 7.5% SDS-PAGE and blotted using anti-Dicer, anti-TRBP 672, anti-Myc and anti-actin antibodies. The lower bands in lanes 1 and 2 correspond to the migration front. B) TRBP2 or TRBP2ΔC4-Dicer interactions by immunoprecipitation. tarbp2-/- cells were transfected with none (lanes 1, 4, 7) or 15 μg of pCMV-Myc-TRBP2 (lanes 2, 5, 8), pCMV-Myc-TRBP2ΔC4 (lanes 3, 6, 9). IP was performed with 1 mg of protein and anti-Myc antibody (lanes 4–6) or anti-Dicer antibody (lanes 7–9). 100 μg of proteins from each lysate (input; lanes 1–3) and the immunoprecipitated complexes (lanes 4–9) were run on a 7.5% SDS-PAGE and blotted using anti-Dicer, anti-Myc and anti-actin antibodies.

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