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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

Fig. 5 ∣. CRBN regulates recombinant and endogenous substrates bearing C-terminal cyclic imides.

Fig. 5 ∣

a, Schematic of HBB(1–cQ132) (HBB native sequence with C-terminal glutarimide at residue 132 instead ofglutamine) and its methylated derivative HBB(1–Me132). b, Quantification of mono-, di- and tri-mono-ubiquitinated HBB band intensity from in vitro ubiquitination of HBB(1–cQ132) or HBB(1–Me132) with K0 ubiquitin. Ordinary one-way ANOVA with Šídák’s multiple comparisons test. Data are mean ± s.d. (n = 3 biologically independent samples). c, Volcano plot of peptide groups bearing C-terminal cyclic imides in HEK293T cells treated without or with 200 μM lenalidomide over 48 h. Upregulated proteins at 1% FDR (red) or 5% FDR (pink); ACTB(96–cN111) peptide (blue). One-way ANOVA with Tukey’s HSD post hoc test. d, Volcano plot of peptide groups bearing C-terminal cyclic imides in MM.1S cells treated with DMSO or 200 μM lenalidomide over 48 h. Upregulated proteins at 1% FDR (red) or 5% FDR (pink); ACTB(96–cN111) peptide (blue). One-way ANOVA with Tukey’s HSD post hoc test. e,f, Quantification of ACTB(96–cN111) and ACTB(96–N111) in HEK293T (e) or MM.1S (f) cells treated with DMSO or 200 μM lenalidomide over 48 h, using SIM. Data are mean ± s.d. (n = 4 biologically independent samples). Unpaired two-tailed t-test; P-values are shown.