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HEK293T cell lysate was incubated with 1 for 10 minutes at room temperature before addition of HA-Ub-VME. This was incubated for 10 additional minutes then quenched by the addition of 4x laemlli buffer and heated at 90 °C for 5 minutes. UCHL1 immunoblot shows a reduction of HA-Ub-UCHL1 formation with increased concentration of 1. HA immunoblot shows two bands decrease as the concentration of 1 is increased, denoted by black arrows.
