Extended Data Figure 2 : CuET is the major anti-cancer metabolite of DSF.

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

Extended Data Figure 2

a, CuET cytotoxicity measured by a colony-formation assay in human cell lines derived from breast, lung, colon and prostate carcinomas. Data are mean ± s.d. of three independent experiments (breast) or presented individually for two independent biological experiments for each cell line (lung, colon and prostate). b, IC50 values from two independent biological experiments documenting differential CuET-induced cytotoxicity across a panel of cancer and non-cancerous cell lines (48 h treatment). c, Analysis of annexin V signal in AMO-1 cells exposed to toxic doses of NMS873 (5 μM, 16 h) or CuET (100 nM, 16 h) and in U2OS cell exposed to toxic doses of NMS873 (10 μM, 16 h) or CuET (1 μM, 16 h). d, Analysis of caspase 3/7 activity in selected cell lines after apoptosis induction by NMS873 (AMO-1: 6 h, 5 μM; Capan1: 16 h, 10 μM; U2OS: 16 h, 10 μM; MDA-MB-231: 24 h, 10 μM) or CuET (AMO-1: 16 h, 100 nM; Capan1: 16 h, 250 nM; U2OS: 16 h, 1 μM; MDA-MB-231: 24 h, 1 μM). e, Absence of cleaved PARP1 after a toxic dose of CuET in U2OS cells, compared to etoposide treatment as a positive control. f, Analysis of cytochrome c (in red) release from mitochondria in U2OS cells during cell death induced by the positive control staurosporin (STS, 1 μM) compared to cell death induced by CuET (1 μM). Blue, DAPI. Scale bar, 10μm. cf, Data are representative of two independent biological experiments.