Dose-Response and Time-Dependence Analysis with the Two Improved ASOs for Splicing of Endogenous <i>SMN1/2</i> Pre-mRNAs

<p>ASOs 07–21 (A) and 34–48 (B) were transfected into HEK293 cells, using starting concentrations from 0 to 20 μM for electroporation. Dose-response curves for ASOs 07–21 and 34–48 were plotted on the right using the data obtained from three independent experiments. Error bars indicate standard deviations. ASOs 07–21 (C) and 34–48 (D) at a concentration of 10 μM were transfected into cells by electroporation. Cells were harvested daily for 5 d to prepare total RNA for splicing analysis as in <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.0050073#pbio-0050073-g002" target="_blank">Figure 2</a>. Exponential-decay curves were plotted on the right using the data obtained from three independent experiments. Error bars indicate standard deviations. The percentage of <i>SMN1</i> and <i>SMN2</i> exon 7 inclusion in each lane is indicated below each autoradiogram.