Case study of PHF20L1. (A) Genomic organization of the cassette exon 6, the complementary sequences R1 and R2, and their respective AONs (AON1 and AON2). (B) The treatment with AON1 or AON2 almost completely suppresses exon 6 skipping; NT (nontreated control); C (control AON); neg (negative control). (C) The scheme of a minigene expressing a fragment of the PHF20L1 gene. Primer locations are indicated by the arrows. (D) Mutagenesis in the minigene. In m1 and m2 mutants, the base pairing between R1 and R2 is disrupted by sequence reversal; in the compensatory double mutant m1m2, it is restored. (E) Exon 6 skipping in m1 and m2 mutants is suppressed, but in m1m2 it returns to that of the WT. Statistically discernible differences at the 1%, and 0.1% significance level are denoted by **, and ***, respectively. Nonsignificant differences are denoted by “ns.”
