Fig. 4
RBM10 modulated alternative splicing of ASPM in CCA. A The number of different alternative splicing events associated with RBM10C761Y mutation in CCA. RI, retained intron; MXE, mutually exclusive exon; A3SS, alternative 3’ splice site; A5SS alternative 5’ splice site; SE, skipped exon. B Significant exon skipping (ES) changes between RBM10 WT and MUT groups (FDR < 0.01, |inclusion level differences| > 0.1). C A flowchart of exploring RBM10 mutation-related ES events. D, E RNA immunoprecipitation using anti-RBM10 or anti-SMN2 antibodies, or igG in different groups. RNA immunoprecipitation (RIP) products were analyzed by RT-PCR. The recovered RNA in each RIP was normalized to its input. F, G Isoforms expression of ASPM in NC, WT, and MUT groups were evaluated at mRNA and protein levels. Green rectangle in (G), CAMSAP CH domain; red rectangle in (G), isoleucine and glutamine domain. H RT-PCR results revealed that the relative expression of ASPM203/ASPM201 was negatively related to RBM10 expression in CCA samples(n = 77) by Pearson rank correlation analysis (r=-0.2989, P = 0.0074). I The effects of RBM10 WT and MUT on ASPM exon18 ES event were examined using minigene splicing reporter assays