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. 2021 Feb 18;118(8):e2007328118. doi: 10.1073/pnas.2007328118

Fig. 2.

Fig. 2.

Effects of compounds on the ubiquitination of CK1α by E3 CRL4CRBN in vitro. (A) Lenalidomide-dependent ubiquitination of CK1α by E3 CRL4CRBN driven by E2 UbcH5c and Cdc34b. (A, Top) The reaction scheme using a procedure as described in SI Appendix, Methods. F-Ub, green; I-Ub-K48R, red. The reaction products were analyzed by both merged fluorescence imaging and immunoblot using an anti-CK1α antibody. (B) KH-4-43 inhibits the ubiquitination of CK1α by CRL4CRBN with Ub-K48R. (C) KH-4-43 inhibits the ubiquitination of CK1α by CRL4CRBN with the wild-type Ub. (D and E) Effects of 33-11 and 33 on the ubiquitination of CK1α by CRL4CRBN. The effects of 33-11 and 33 shown in D and E were determined using assays similar to B and C, respectively.