Effect of siRNA Treatment on WT and Mutant Allele Frequency Assessed by qRT-PCR.

<p>A) Schematic diagram depicting the allele-specific PCR primer design used to determine fractional WT and mutant allele expression. The IT (<i>top</i>) and YS (<i>bottom</i>) primer sets amplify 147 and 159 bp PCR products, respectively. Each primer set includes an upstream primer common for both the WT and mutant alleles and a downstream primer designed to specifically anneal to only the WT or mutant template. B) The result of normalized ΔΔCt analysis comparing the fractional contribution of mutant to WT allele frequency (mutant/WT) such that an equal expression is reflected in a value of 1.0 and an increase in fractional WT allele frequency reflected in values <1.0. IT/+ mice were treated for two weeks with either SCR or IT-Dc siRNA. YS/+ mice were treated for two weeks with either SCR or YS-Dc siRNA. A total of n = 4 FDB muscles were used for all conditions. ^* p<0.01.