Fig. 5.

Cotreatment with anion-channel blockers and transient knock-down of LRRC8A abolish Cisplatin-induced but not TNFα and Hyperosmotic-induced Caspase-3 activation in wild-type A2780 cells. A2780WT and A2780CisR cell lysates were used for Caspase-3 activity assay using a commercial kit (see experimental procedures). A: A2780CisR cells were exposed to 10 μM Cisplatin for 18 h and A2780WT cells were likewise treated with Cisplatin in the absence or presence of 100 μM NS3728 or 400 μM DIDS. Data represent 6–8 (A2780WT) and 4 (A2780CisR) individual sets of experiments ± SE. * and #: Significantly increased by Cisplatin compared with the respective control and significantly reduced compared with Cisplatin treatment in the absence of the inhibitor, respectively (ANOVA, Fisher LSD method). B: A2780WT were treated with either 25 nM scramble siRNA or LRRC8A siRNA for 30 h and subsequently exposed to 10 μM Cisplatin for another 18 h. Data represent 4 individual experiments ± SE. * and #: Significantly increased by Cisplatin compared with the respective control and significantly reduced compared with Cisplatin treatment in Scramble siRNA-treated cells (Student's t-test). C: A2780CisR cells were exposed to 20 nM TNFα for 18 h. A2780WT cells were likewise treated with TNFα in the absence or presence of 100 μM NS3728 or 400 μM DIDS. Data represent 6–8 (WT) and 4 (CisR) individual experiments ± SE. *Significantly increased compared with the respective control (ANOVA, Fisher LSD method). D: A2780WT cells were exposed to isotonic or hypertonic NaCl for 4.5 h in the absence or presence of 100 μM NS3728 or 400 μM DIDS. Data represent 3 individual experiments ± SE. *Significantly increased compared with isotonic control (ANOVA, Fisher LSD method).