Figure 5.

The effect of two over-abundant overlaps in a minigene reporting system. Each tested overlap was inserted into the second exon of a reporting minigene comprised of four exons. (A) Illustration of an overlap comprised of two spicing regulatory sequences. Seq1 is in light-gray, seq2 is in black, and the overlapping sequence is in dark-gray. For each overlap, which is comprised of two splicing regulatory motifs (seq1 and seq2), we introduced two mutations: (B) mutation to eliminate seq1 leaving seq2 intact, and (C) mutation to eliminate seq2 leaving seq1 intact. (D) Insertion of both mutations simultaneously. (E) Splicing patterns of each of the two overlaps (overlap I: lanes 1–4; and overlap II: lanes 5–8) is depicted. For each overlap, the splicing pattern in its presence, after the elimination of seq1, the elimination of seq2, or the elimination of both is depicted, respectively. The upper bands indicate exon inclusion, while the lower bands indicate exon skipping; the exact inclusion level is indicated above each lane.