Figure 5

24S-OHC induced apoptosis and necroptosis in Jurkat cells. (a) Jurkat cells were pretreated with 20 μM ZVAD for 1 h and were then treated with 50 μM 24S-OHC for 24 h. Cell viability was measured by WST-8 assay. N.S., not significant. **P<0.01, when compared with cells without ZVAD and 24S-OHC; ^††P<0.01, when compared with ZVAD-treated cells. (b) Jurkat cells were pretreated with 20 μM ZVAD for 1 h and were then incubated with 50 μM 24S-OHC for 6 h. As a control, cells were treated with 100 ng/ml CHX and 100 μg/ml TNFα. Whole cell lysates were immunoblotted with appropriate antibodies as indicated. (c) Jurkat cells were pretreated with 20 μM ZVAD and 100 μM Nec-1 for 1 h and were then treated with 50 μM 24S-OHC for 16 h. Cell viability was measured by flow cytometry using propidium iodide. *P<0.05. (d) Caspase-8-deficient Jurkat cells were pretreated with 100 μM Nec-1 for 1 h and were then treated with 50 μM 24S-OHC for 16 h. Cell viability was measured by flow cytometry using propidium iodide. **P<0.01. (e) Jurkat cells were transfected with RIPK3 (siRIPK3) or negative control (NC) siRNA oligo for 72 h. The cells were pretreated with 20 μM ZVAD for 1 h and were then treated with 50 μM 24S-OHC for 24 h. Cell viability was measured by WST-8 assay. **P<0.01 when compared with NC siRNA+24S-OHC+ZVAD