Fig. 3.

TAKI-induced cytotoxicity is dependent on mitochondrial ROS. (a, b) BMDM were treated with TAKI (100 nM), LPS (100 ng/ml), or both for indicated time periods. (c, d) BMDM were pre-treated with Nec-1 (10 μM) or zVAD (20 μM) for 30 min before the addition of TAKI, LPS or both for 6 h. The cytosolic and mitochondrial ROS production were determined by DCFH-DA (a, c) and MitoSOX (b, d), respectively, and we used the levels under vehicle treatment as the control. (e, f) BMDM were pre-treated with NAC (5 mM) or BHA (150 μM) for 30 min followed by TAKI and/or LPS stimulation. After 6 h incubation, LDH release was determined (e) and after 4 h drug treatment, Annexin V/PI staining (f) was determined. *p <0.05, indicating significant effects of TAKI or LPS alone. #p <0.05, indicating the significant potentiation of TAKI-induced responses by LPS. **p <0.05, indicating significant inhibitory effects of Nec-1, NAC and BHA on the responses induced by TAKI, either in the absence or presence of LPS