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. Author manuscript; available in PMC: 2020 Oct 3.
Published in final edited form as: Mol Cell. 2019 Aug 29;76(1):82–95.e7. doi: 10.1016/j.molcel.2019.07.017

Figure 6. Other SF3B1 Mutations Also Weaken Interaction with SUGP1 in the Spliceosome.

Figure 6.

(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).