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. 2013 Jul 5;3:2142. doi: 10.1038/srep02142

Figure 1. Construction of the Clcn1-L reporter minigene and effect of triplet repeat expansion on the reporter vector Clcn1-L.

Figure 1

(a) Schematic structure of the Clcn1-L minigene reporter. A genomic segment of mouse Clcn1 containing exons 6 to 7 (including the intron) was sub-cloned downstream of EGFP in the pEGFP-C1 plasmid. Firefly luciferase was inserted in-frame with a correct Clcn1 splicing pattern. (b) Schematic of how Clcn1-L functions. Exon 7A exclusion results in luciferase expression to detect correct Clcn1 splicing. (c) Inclusion of Clcn1 exon 7A increased upon expression of the expanded CUG repeat. (d) Bar charts show the quantified percentages of exon 7A inclusion (mean + SEM, n = 4). (e) Luciferase analysis showed that relative luciferase activity decreased upon expression of the expanded CUG repeat (mean + SEM, n = 3). The gel image was cropped around the region of interest and the samples (n = 4) were resolved in the same gel. Statistical significances were determined using t-tests (*p < 0.05, ***p < 0.001).