Knockdown of SR and hnRNP proteins modulates endogenous <i>SMN2</i> exon 7 inclusion.

<p>(<b>A</b>) Semi-quantitative radiolabelled RT-PCR of endogenous <i>SMN2</i> mRNA following RNAi-mediated knockdown of individual SR or hnRNP proteins in HeLa cells. Reaction products were digested with DdeI. Digestion products are labeled. (<b>B</b>) Graphs show the quantification of <i>SMN2</i> exon 7 inclusion: (included/skipped). Asterisks indicate a statistically significant change in exon 7 inclusion, *p≤0.05, **p≤0.01 and ***p≤0.001, unpaired Student’s T-tests. (<b>C</b>) Quantification of RT-PCR confirmed effective, selective knockdown of SR proteins and hnRNPs. The dashed line represents the level of control expression. In all cases, error bars represent SEM. For SRSF1 and SRSF2 n = 3; SRSF4, SRSF6, hnRNP A2/B1, ELAVL1 and hnRNP D n = 4; SRSF7, SRSF10, and hnRNP U n = 5; SRSF9, hnRNP C, hnRNP H1 and hnRNP F n = 6, SRSF5 n = 7, SRSF3 and SRSF11 n = 8; Control cells were transfected with a scrambled control siRNA. (<b>D</b>) Immunoblot analysis of indicated endogenous protein from lysates of cells treated with siRNAs targeting the indicated SR protein or a non-specific control siRNA. β-actin is a loading control.