Figure 5.

TRBP1 and TRBP2, but not TRBPsΔC4 rescue the RNAi pathway in tarbp2^-/- cells. A) TRBP1 and TRBP2 rescue the RNAi pathway in tarbp2^-/-cells. tarbp2^-/- cells were transfected with none or 1 μg of EGFP and 4 μg of non-silencing (ns) or anti-EGFP (gfp) shRNA as indicated, and 1 μg of pcDNA3 (left panels), pcDNA3-TRBP1 (middle panels) or pcDNA3-TRBP2 (right panels). (Top panels) EGFP knock-down was assessed by fluorescence, and average percentages of EGFP knockdown were calculated from luminosity intensity values of four fields per condition. Representative photos are shown. (Bottom panels) Cell extracts were analyzed by immunoblotting against EGFP and actin. Percent decrease of EGFP signal was calculated by densitometry analysis. B) TRBP1ΔC4 and TRBP2ΔC4 do not rescue the RNAi pathway in tarbp2^-/- cells. tarbp2^-/- cells were transfected with 1 μg of EGFP and 4 μg of non-silencing (ns) or anti-EGFP (gfp) shRNA as indicated, and 1 μg of either pCMV, pCMV-Myc-TRBP1, pCMV-Myc-TRBP1ΔC4, pCMV-Myc-TRBP2 or pCMV-Myc-TRBP2ΔC4 as indicated. (Top panels) EGFP knock-down was assessed by fluorescence, and average percentages of EGFP knockdown were calculated from luminosity intensity values of four fields per condition. Representative photos are shown. (Bottom panels) Cell extracts were analyzed by immunoblotting against EGFP and actin. Percent decrease of EGFP signal was calculated by densitometry analysis.