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. Author manuscript; available in PMC: 2023 Oct 6.
Published in final edited form as: Nat Cell Biol. 2023 Apr 6;25(4):528–539. doi: 10.1038/s41556-023-01109-9

Extended Data Fig. 7. DNMT3A associates with components of the spliceosome.

Extended Data Fig. 7

(A) Western blot validation of biotin-tagged DNMT3A after induction with 2ug/mL doxycycline in mouse ES cells (left) and immunoprecipitation with streptavidin coated magnetic beads (right). DNMT3A (at 130 kDa) is labeled. Red box indicates fractions of cytoplasmic and nuclear cell lysates used for mass spectrometry. (B) Table showing top splicing associated factors enriched in the co-immunoprecipitation and mass spectrometry analysis of biotin-tagged DNMT3A in mouse ES cells. (C) BiFC analysis of DNMT3A-interacting spliceosome components after lentiviral expression of human DNMT3A variants (n = 4 biologically independent samples and 2 independent experiments, mean values +/− SD, unpaired student’s t-test with Welch’s correction, mean values +/− SD, p < 0.05 = ** ). Bar graph depicting analysis of Bimolecular Florescence complementation (BiFC) where fluorescence signal is recorded when each component of the spliceosome (prey) was introduced to (D) human DNMT3A, (E) human DNMT3A R882H, (F) human DNMT3B (bait). 2 biologically independent experiments. Data shown represents mean values +/− SD.