Fig. 1. Co-selected mutations in VCP confers resistance to CB-5083.

. a Presence of both wildtype VCP alleles results in wildtype VCP protein hexamer, which can be inhibited by CB-5083. Similarly, presence of one mutant allele (E470K or E470D) result in VCP protein hexamers with mixed VCP proteins (wildtype and mutant). However, inhibition of wildtype protein in the complex upon CB-5083 treatment is sufficient for cytotoxicity. Loss of wildtype copy is required for resistance towards CB-5083. b Collective mutations observed upon CB-5083 treatment can occur via three scenarios. Scenario 1 outlines the chance of two different mutations occurring simultaneously; whereas scenario 2 and 3 outline the occurrence of serial mutations whereby mutations at D1–D2 linker is then followed by mutations at D2 domain or vice versa