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. Author manuscript; available in PMC: 2023 Jul 3.
Published in final edited form as: Nature. 2022 Oct 19;610(7933):775–782. doi: 10.1038/s41586-022-05333-5

Extended Data Fig. 3 ∣. Co-immunoprecipitation and photo-affinity displacement assay with dipeptide degraders and ligands.

Extended Data Fig. 3 ∣

(a) Co-immunoprecipitation of endogenous BRD4 from HEK-CRBN in cells (left) or in lysates (right) after 2 h treatment with 25 μM or 1 μM of the indicated degrader. Western blot data are representative of 2 independent replicates. (b) Photo-affinity labeling displacement assay using photo-lenalidomide (pLEN) to visualize in-gel fluorescence imaging of CRBN/DDB1. Comparisons were performed using unpaired two-tailed t-tests. ns = not significant, * = p < 0.05, ** = p < 0.01, *** = p < 0.001, **** = p < 0.0001. Data are presented as mean ± SD (n = 3 biologically independent samples). (c) Structure of photo-lenalidomide (pLEN) and schematic of photo-affinity labeling displacement assay using photo-lenalidomide (pLEN) to visualize in-gel fluorescence imaging of CRBN/DDB1. (d) Corresponding gel images after treatment of CRBN/DDB1 with 1 μM pLEN and 100 μM of the indicated competitor for 30 min prior to photo-affinity labeling and in-gel fluorescence imaging. Data shown include 3 biologically independent replicates. For uncropped western blot and gel images, see Supplementary Fig. 9.