Figure 10.

Mechanistic characterization of VZ185 mode of action. (A) VZ185 activity is proteasome and CRL2^VHL-dependent. RI-1 cells were treated in two replicates with MG132, MLN4924, VH298, and cisVZ185 in the presence of absence of VZ185 (100 nM) at desired time points before harvesting. (B) NanoBRET experiments in HEK293 cells to monitor ubiquitination of HiBiT endogenously tagged BRD9 (purple) or BRD7 (green) using monoclonal-Ub primary and Alexa594 secondary antibodies at times indicated after treatment with 1 μM VZ185. Error bars expressed as SEM of n = 3 experiments.