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. Author manuscript; available in PMC: 2015 Oct 1.
Published in final edited form as: Leukemia. 2014 Oct 14;29(4):909–917. doi: 10.1038/leu.2014.303

Figure 5. Recurrent missense mutations in U2AF1 have specific effects on alternative splicing.

Figure 5

(a) U2AF1 mRNA expression was increased >10-fold in cells transfected with WT or mutant alleles, compared to MIG control. Error bars (too small to visualize) represent SD of 3 technical replicates; repeated in 3 biological replicates for each construct with similar results. (b) Exogenous Flag-tagged U2AF1 protein abundance of the different mutants in 293T cells compared with MIG empty vector control. Numbers beneath the blot are the sum of U2AF1- Flag and endogenous U2AF1 values in each case, normalized to MIG, obtained by densitometry analysis. (c) The U2AF1 S34 mutants (S34F>S34Y) increased exon skipping in the GH1 minigene. Exon skipping was decreased by the Q157R mutant, and unaffected with an allele containing both mutations (S34F/Q157R), compared to WT U2AF1. (d) Similar mutation- specific effects were seen with cryptic splicing of the FMR1 minigene. (e) Exon skipping of endogenous DEK was increased by the U2AF1 mutants relative to WT in 293T cells but decreased by the S34Y, S34F/Q157R and Q157R mutants, compared to the S34F mutant. The splicing ratio (b/total) = expression of isoform b/(the sum of the expression of isoforms a and b). The mean +/− SD of 3 replicates are shown; repeated in 3 separate experiments with similar results. *P<0.05, **P<0.01, ***P<0.001.