Figure 6. Other SF3B1 Mutations Also Weaken Interaction with SUGP1 in the Spliceosome.
(A and B) HEK293T cells in 10-cm plates were transfected with empty vector plasmid (Vec), or expression plasmid for His6-FLAG-tagged WT, K700E, E622D, R625C, H662Q, or K666N SF3B1, followed by affinity purification using the small-scale protocol. The SF3B1-associated proteins were resolved by SDS-PAGE, followed by silver staining (A) or western blotting (B). Marker, Precision Plus Protein marker (Bio-Rad).
(C and D) HEK293T cells in 6-well plates were transfected with empty vector plasmid (Vec), or expression plasmid for HA-tagged WT, K700E, E622D, R625C, H662Q, or K666N SF3B1, followed by RT-PCR of the cryptic 3′ss (open arrowheads) and canonical 3′ss (solid arrowheads) of GCC2 and KANSL3 (C) or western blotting (D).
