Figure 5: MBNL1 knockdown exacerbates DM1-like splicing shift induced by compound 1 in normal human fibroblasts.
From: Induction and reversal of myotonic dystrophy type 1 pre-mRNA splicing defects by small molecules
(a) Representative RT–PCR assays showing splicing changes in MBNL1-dependent exons of four genes in normal fibroblasts treated with siRNA against MBNL1 (siMBNL1) and increasing amounts of 1 (n=2). Mock controls are lipofectamine-treated samples. (b) Quantification of alternative splicing shift towards the DM1-like phenotype (% of alternative exon inclusion) in four MBNL1-regulated exons (MBNL1, MBNL2, NCOR2 and NFIX) upon combined treatment of normal human fibroblasts with an siRNA against MBNL1 and increasing concentrations of 1 (orange bars; n=2). For comparison, splicing shifts in siMBNL1-treated DM1 affected 500CUG fibroblasts are shown (blue bars; n=2). Mock controls represent splicing changes in lipofectamine-treated cells (green bars—normal fibroblasts; violet bars—500CUG fibroblast; n=2). ‘*’ indicates P≤0.05; ‘**’ indicates P≤0.01; and ‘***’ indicates P≤0.001 as determined by a two-tailed Student’s t-test.
