Extended Data Figure 3 : CuET-induced proteasome inhibition-like response is not due to proteasome inhibition.

From: Alcohol-abuse drug disulfiram targets cancer via p97 segregase adaptor NPL4

Extended Data Figure 3

a, Kinetics of poly-Ub protein accumulation in U2OS cells treated with CuET or the proteasome inhibitor BTZ. b, CuET treatment (1.5 h) induces rapid deubiquitylation of ubiquitylated histone H2A (uH2A) similarly to proteasome inhibitors BTZ or MG132 in U2OS cells. c, CuET treatment (1.5 h) induces rapid cytoplasmic accumulation of poly-ubiquitylated proteins (FK2 antibody staining) in U2OS cells, similar to BTZ and MG132 treatment. Scale bar, 10μm. d, e, 20S proteasome activity is not inhibited by CuET as examined in live MDA-MB-231 cells (d) or in lysates from MDA-MB-231 cells (e). Data are mean ± s.d. of four independent experiments. f, CuET treatment (1 μM, 6 h) does not cause accumulation of p53 in the presence of dicoumarol (300 μM) in MCF7 cells. g, In vitro 26S proteasome function measured as RPN11 deubiquitylation activity, is not inhibited by CuET; 1,10-phenanthroline (1,10-OPT) served as a positive control. Data are representative of two (ac, f) or three (g) independent experiments.