Figure 3.

Minigene splicing assay confirmed the pathogenicity of the mutations. (A) HEK293T cells were transfected with wild-type or mutant minigene plasmids. The wild-type minigene expressed both fluorescent proteins, whereas both mutants had limited EGFP expression, indicative of aberrant gene splicing. (B) Flow cytometry analysis of transfected HEK293T cells showed significant quantitative differences in the percentage of cells expressing EGFP. Tukey box and whisker plot, where center line represents median, box limits represent interquartile ranges, and whiskers represent minimum to maximum data ranges; black square, outlier; the mean values for the mutants overlap the lower line in the boxes. *P < 0.05; ****P < 0.0001; n.s., not significant.