Fig. 3: The turnover of TOP2β upon VM-26 treatment is dependent on the β-TrCP degron motif of TOP2β. | Oncogenesis

Fig. 3: The turnover of TOP2β upon VM-26 treatment is dependent on the β-TrCP degron motif of TOP2β.

From: SCFβ-TrCP-mediated degradation of TOP2β promotes cancer cell survival in response to chemotherapeutic drugs targeting topoisomerase II

Fig. 3

a Diagram of mutants of two potential β-TrCP degron motifs in TOP2β. b TOP2β S1130A and S1134A mutants, but not the S1316A mutant, have longer protein half-lives. HEK293 cells transfected with wild-type or indicated mutants of FLAG-TOP2β were treated with CHX and VM-26 for the indicated time periods, and then, IB was undertaken using the indicated Abs (left). Densitometry quantification was performed with ImageJ, and the decay curves are shown (right). c Reduction in β-TrCP-TOP2β binding by degron site mutations. HEK293 cells transfected with the indicated plasmids were treated with MG132 and VM-26 for 5 h, and then, IP was conducted with anti-FLAG beads (top), and direct IB was undertaken with the indicated Abs (bottom). d Reduction in TOP2β ubiquitination by degron site mutations. HEK293 cells transfected with the indicated plasmids were treated with MG132 and VM-26 for 5 h, and then, IP was conducted using anti-HA beads (top), and direct IB was undertaken using the indicated Abs (bottom).

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