Fig 6. Short term disulfiram/copper treatment causes irreversible cell damage, associated with cell detachment and actin fiber depolymerization.

A) OVCAR3 cells grown on glass cover slips were treated for 4 and 8 h with 1 μM disulfiram/copper, fixed with methanol, and incubated with fluorescent Bodipy® FL phallacidin to visualize F-actin fibers. B) OVCAR3 and SKOV3 cells grown 24-well cell culture dishes (1 × 104 cells/well) were treated as in (A) for 4 h and 8 h with 1 μM disulfiram/copper, collected by trypsinization in 500 μl trypsin/EDTA solution (Biochrom, Berlin, Germany), and reseeded at a concentration of 500 cells/well in 6-well cell culture plates. After further 10 days of incubation, cell clones were fixed with methanol and stained with Coomassie brilliant blue.