Figure 4: BCLAF1 was a conserved and direct target of SRSF10.

(a) Diagram of exon5a (5a1, 5a2 and 5a3) and exon5 RNA, and gel-shift assays. Indicated ³²P-labelled RNAs were incubated with increasing amounts of recombinant His-SRSF10 (100 and 300 ng) and complexes were resolved by non-denaturing PAGE. (b) 293T cells were transiently transfected with HA-tagged SRSF10, SRSF1, SRSF2 or empty-vector controls. Western blotting analysis was performed to confirm the expression of each protein by using indicated antibodies. (c) In vivo ultraviolet cross-linking and immunoprecipitation (CLIP) from 293T cells described in b with primer pairs complementary to human BCLAF1 exon5a1, 5a2 and 5a3 or exon5 by RT–PCR. (d) RNAs were extracted from SRSF10 siRNA (si-SRSF10)- and control siRNA (si-NC) -transfected cells. RT–PCR was performed as described in Fig. 1b. (e) Schematic diagram of RNA-seq reads coverage over BCLAF1 exons 4–6 between si-NC and si-SRSF10 cells. (f) Whole-cell lysates were prepared from cells described in d and analysed by western blotting using indicated antibodies.