Number and severity of splice defects increase when MBNL1 is silenced incrementally from ∼79% to ∼98%.

<p>SkMC were transfected with siRNAs directed against MBNL1 and cell samples on each subsequent day post-siRNA transfection for a period of 5 days, were divided into 4 aliquots where one aliquot was used to measure MBNL1 levels and total RNA was extracted from each of the three other aliquots. Scrambled siRNA transfected samples were harvested on Day 5, the last time point of the experiment. <b>(A)</b> Total protein (10 µg) was analyzed by western blot to measure the silencing achieved for MBNL1 at 24 h intervals for 5 days. Blots were probed for GAPDH as an internal control. <b>(B)</b> Synthesized cDNAs were subjected to PCR analysis to study RNA splicing as indicated with <i>GAPDH</i> RNA as an internal control. In each case the levels of exon inclusion obtained in the experiment shown are indicated. <b>(C)</b> The results of RNA splicing as a function of MBNL1 levels in SkMC are tabulated.