Figure 3.

Intronic deletion analysis of Clcn1. (A) Structure of Clcn1 deletion mutants. Deletion series of the Clcn1 minigene were generated by PCR-mediated mutagenesis. FL corresponds to the full-length Clcn1 minigene covering exons 6–7 and is the same construct as that used in Figures 1 and 2. The positions of nucleotides at the termini of exons and the junction of deleted regions are indicated. (B) Splicing analysis of Clcn1 deletion mutants in COS-7 cells. Cellular-splicing assays were performed using deletion mutants Δ1–Δ9 and MBNL1. Lanes ‘V’ and ‘M’ indicate the co-expression of each deletion minigene with empty vector and MBNL1, respectively. Splicing patterns were detected by RT–PCR, as in Figure 1B.