Q157P, Q157R, and Q157Rdel have distinct 3′ splice site sequence preferences. (A) Identification of U2AF35 target exons responding to Q157P, Q157R, and Q157Rdel variant expression in RNA sequencing data. Cassette exons with increased skipping upon U2AF35 knockdown and rescue by wt U2AF35 expression were defined as U2AF35 target exons. Overlap of Q157P, Q157R, and Q157Rdel responsive U2AF35 target exons is shown together with the 3′ splice site consensus motifs of unique Q157P, Q157R, and Q157Rdel targets. (B) Splicing of BCL2L12 alternative exon 3 in response to U2AF35 knockdown and rescue by U2AF35 variants. Splicing of endogenous BCL2L12 (left, n = 3) was compared to the wild-type and mutated minigenes of BCL2L12 containing exons 2, 3, and 4. Minigenes were transfected in duplicates in two independent experiments.
