Fig. S3.

Additional validation of potent sense strand inhibition from a single plasmid coexpressing shRNA and TuD. (A) Confirmation that both perfect and imperfect sense strand targets can be rescued by TuD coexpression as determined by cotransfection of HEK293T cells with the indicated shRNA/TuD plasmids or no-shRNA control and dual luciferase reporters containing the shown binding sites for the shRNA sense or antisense strand. (B–D) Validation that sense strand targets can be rescued by TuD coexpression using a fluorescent reporter system, as observed by fluorescence microscopy (B), quantitative FACS analysis (C), and mRNA Northern blot analysis (D) of HEK293T cells transfected with the indicated plasmids. In D, “Quant.” (quantification) indicates relative egfp band intensities normalized to mCherry intensities. **P < 0.01; two-sided t-test. Error bars in A and C represent SDs of at least three independent experiments.