Fig. 2
Effects of BV-6 treatment on KGN. The SMAC mimetic BV-6 was used in different cell viability assays to explore the effects on KGN. Different BV-6 concentrations (0.05–100 μM) were tested. (a) Live cell images of KGN (passages > 80) treated with BV-6 (0.1, 1, 10 and 50 μM) for 24 h. Corresponding solvent controls are shown in insets. At 0.1 and 1 μM no indications for cell death were evident. At 10 μM detached cells were visible. At 50 μM all cells detached. Scale bar indicates 50 μm. (b, left graph) Subsequent cell counting analysis of KGN (passages > 80) treated with BV-6 (0.05–100 μM) revealed an EC50 of 7.4–8.2 μM after nonlinear regression analysis. (n = 3, bars indicate SEM). (b, right graph) ATP assay with BV-6 (0.1–100 μM) confirmed the cell counting experiment. Nonlinear regression analysis revealed an EC50 ranging from 7.2 to 9.7 μM. (n = 4, error bars indicate SEM). (c) To examine a possible involvement of the passage number, KGN from early passages (< 8) were stimulated with BV-6 (0.05–100 μM) for 24 h and then counted. Nonlinear regression analysis revealed an EC50 of 8.1 - 8.6 μM. (d) time dependence was evaluated by confluency measurement (20 min intervals) of BV-6 (EC50, 8 μM) treated KGN. Data were normalized to the solvent control. Decline in confluency started after 12 h and reached 0.58 (control =1.0) after 24 h. (n = 3, error bars indicate SEM, rel. = relative)