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Priming with UbcH5c∼Ub reverses inhibitory effects by KH-4-43 on the ubiquitination of CK1α. The prior reaction of CK1α-CRL4CRBN-lenalidomide with UbcH5c∼Ub caused resistance to KH-4-43 inhibition. The reaction scheme is shown on the top. The detailed procedure is described under “Experimental Procedure.” Species∗ most likely represents the auto-phosphorylated form of CK1α (27). The substrate levels after reactions were quantified and shown. This experiment was repeated once.
