Figure 2.
Blocking endogenous DUB activity with NEM and TCEP. Cells were harvested in the absence or presence of TCEP, and with the addition different NEM concentrations followed by blocking the DUB activity for 2 h on ice. Then the excess of NEM was quenched with 20 mM DTT for 2 h on ice and the efficiency of DUB inactivation was tested by incubating the lysates overnight (O/N) at 37 °C. (B) Cells harvested without TCEP and (C) cells harvested with 0.5 mM TCEP.