Skip to main content
. 2019 Apr 22;47(11):5922–5935. doi: 10.1093/nar/gkz259

Figure 5.

Figure 5.

Increased structural stability around the normal splice site induces usage of the alternative splice site without the C1824U point mutation. (A, B) Design of the WT/Norm-SS-Closed-1 and WT/Norm-SS-Closed-2 mutants, generated from the WT minigene construct. The normal 5′ SS sequence is highlighted by a blue line. Ins. A: Sites at which adenines are inserted. (C) RT-PCR analysis for LMNA/Δ150 expression levels of each mutant. 293T cells were transfected with 1 μg of the indicated plasmids. RNA was harvested at 72 h post transfection. DsRed expression served as an internal loading control. (D) Quantification of mRNA isoforms expressed as relative usage of the normal versus alternative 5′ SS. Data are expressed as mean ± s.d. from three independent experiments. UT: untransfected; EV: empty vector; WT: LMNA wild-type minigene construct; MUT: LMNA C1824U mutant minigene construct; NC: negative control (PCR reaction mix without cDNA template).