Figure 8

Blocking NFκB-signalling sensitises NKT cells to TNF-α-induced apoptosis. Thymocytes were treated with the IKK inhibitor BMS345541 or DMSO. After an hour, cells were washed and stimulated with indicated amount of TNF-α. After 14 hours, cells were washed and treated with fluorescently-tagged, active caspase 3 (A,D), caspase 8 (B,E) or caspase 9 (C,F) substrates separately. After four hours, NKT cells were identified by staining with anti-CD3ε and CD1d-tetramer and dying/dead cells were identified by staining with 7-AAD. Representative dot plots and quantification of caspase⁺7-AAD⁺ cells within electronically gated CD8^lo thymocytes are shown. Data are mean + sem from over three independent experiments. ns, not significant (P > 0.05), **P ≤ 0.001, ***P ≤ 0.0001.