Fig. 7
Mutant SF3B1-associated alternative 3′ splice sites. a, b Mutant SF3B1-associated alternative 3′ splice sites in ABCB7 (a) and PPOX (b). RNA-seq coverage is shown for samples with mutated and wild-type SF3B1, as well as for those with and without CHX treatment only in a. The bottom figure in a illustrates an RNA sequence, a spliced transcript, and the corresponding amino acids near the 3´ splice site of exon nine of ABCB7. c, d RT-PCR of the differentially spliced sites of ABCB7 (c) and PPOX (d). RT-PCR was performed for primary MDS samples and CRISPR cell lines with or without the SF3B1K700E mutation. Clones #1 to #3 carried a heterozygous SF3B1K700E allele, while clones #4 to #6 had only wild-type alleles. Samples without reverse transcription (RT) were used as a negative control. Samples treated with CHX are also shown in c. Canonical and abnormal transcripts are indicated by blue and red triangles, respectively. e Immunoblots of CRISPR cell lines with or without the SF3B1K700E mutation. Lysates were processed for immunoblotting with antibodies against denoted proteins. f, g Relative protein levels of ABCB7 (f) and PPOX (g). The dot plots show protein levels in three independent clones. Statistical significance was determined by the unequal variance t-test; t-values were 6.9 (f) and 4.3 (g)
