Figure 2

cis-UCA induces both apoptotic and necrotic cell death in 5637 bladder carcinoma cells. The cells were treated with indicated concentrations of cis-UCA in buffer solutions adjusted to pH 7.4 or pH 6.5 for 2 h and let to recover in the absence of cis-UCA for 20 h. The cell culture supernatants and cell lysates were analysed for formation of cytoplasmic histone-associated DNA fragments with a photometric enzyme-immunoassay (A.). Alternatively, the cells treated with cis-UCA or PIPES (corresponding to 2% cis-UCA equimolar concentration) at pH 6.5 were collected and labelled with TMRM for mitochondrial membrane potential (B.), analysed for caspase-3 activation (C.) or labelled with PI for cell permeabilization as a measure of non-apoptotic cell death (D.). The samples were analysed by flow cytometry (Mean ± SEM; n = 3; Student's t-test * p < 0.05, ** p < 0.01, *** p < 0.001). The pictures are representative of two independent experiments.