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. Author manuscript; available in PMC: 2021 Feb 12.
Published in final edited form as: Nat Struct Mol Biol. 2021 Jan 21;28(2):190–201. doi: 10.1038/s41594-020-00550-w

Fig. 6. An artificial IDR-containing KDM5A fusion protein phenocopies NUP98-fusion-induced changes in the condensome.

Fig. 6

a, Schematic illustration of artificial (art) KDM5A fusion proteins. Thirteen triple repeats of phenylalanine-glycine (FG) or alanine-alanine (AA), connected by 18-amino-acid linkers, were fused to the C-terminal part of KDM5A found in patients with NUP98–KDM5A-driven AML. b, Western blot analysis of HEK293T cells transfected with mock, artFG–KDM5A or artAA–KDM5A. Fusion proteins were detected with anti-HA antibodies, and tubulin was used as a loading control. Blot is representative of three independent experiments. Uncropped images are available in Supplementary Fig. 1. c, Live cell imaging of HEK293T cells expressing GFP-tagged variants of artFG–KDM5A and artAA–KDM5A. Fusion proteins are shown in white, and Hoechst staining is shown in green. Scale bar, 5μm. Images are representative of six independent experiments. d, Schematic illustration of biCon-MS for HL-60 cells expressing artFG–KDM5A or artAA–KDM5A. MS analysis was performed in two biological and two technical replicates. e, Venn diagram of proteins enriched in 33μM precipitates compared to 11 μM precipitates for artAA–KDM5A (Extended Data Fig. 6a) and artFG–KDM5A (Extended Data Fig. 6b). f, Reactome FI clustering for 237 proteins uniquely enriched in artFG–KDM5A compared to artAA–KDM5A. The three most significant nuclear clusters are shown with String db interactions (cutoff 0.4) for individual proteins. Gray border thickness indicates PScores for individual proteins.