FIGURE 1

Sequence specificity and efficacy of K6a 3'-UTR siRNAs. (a) Alignments of portions of the human K6a and K6b 3'-UTR cDNA sequences, showing that the target sequences of the four siRNAs (bold type) are located where several insertion/deletion and single-base substitutions occur between K6a and K6b. (b) Normalized K6a-EYFP expression measured by FACS of 293FT cells transiently transfected with K6a-EYFP and the lead K6a siRNAs (K6a-S1, -S2, -S3, and -S4). A potent EGFP siRNA (which also targets EYFP equally well) was used as a positive control. A non-specific control siRNA (NSC4) did not significantly affect expression, even at high concentrations, whereas all four K6a inhibitors worked as well or better than the EGFP control inhibitor at low sub-nanomolar concentrations. Bars on the graph represent the meanSE. (c) Visual confirmation of K6a siRNA efficacy by wide-field imaging of 293FT cells transiently transfected with K6a-EYFP and siRNAs over a range of inhibitor concentrations.