Figure 1. Citronellol induced necroptosis in NCI-H1299 cells. (A) NCI-H1299 cells treated with 49.74 μg/ml (24 h IC50) citronellol for 24, 48 and 72 h. Cells were stained with FITC-conjugated Annexin V-PI and measured by flow cytometry. Control: 24 h + medium: Ethanol 0.5%, 24 h + ethanol 0.5% + medium. (B) NCI-H1299 cells were treated with 49.74 μg/ml citronellol for 6, 12, 18 and 24 h. Cells lysates were immunoblotted with TNF-α, phosphoRIP3, phosphoRIP1 antibodies, and β-actin was used as the loading control. Numbers below represented the relative protein quantitation. C: control, 24 h + medium; E: 24 h + Ethanol 0.5% + medium. (C) Western blot data. (D) Western blot data for pro/active caspase- 3, pro/active caspase8 and pro/cleaved PARP. (E) Western blot data. Experiments were repeated at least three times.