Fig. 2

EBV-LMP1 plays a role in suppression of T/S/Z-induced necroptosis. a, b CNE1 and CNE1-LMP1 cells were treated with T/S/Z, T/S/Z + Nec-1, T/S/Z + GSK’872, Nec-1, GSK’872, or DMSO solvent control for 72 h. At the end of treatment, cell viability, and cell survival were determined by MTS and trypan blue exclusion assay. Data were presented as mean ± S.E.M. (n = 3). P values are by Student’s t test. T TNF-α, 100ng/ml; S Smac mimetic, 5 μM; Z z-VAD-fmk, 20 μM; Nec-1 Necrostatin-1, 40 μM; GSK’872: 5 μM. c CNE1 and CNE1-LMP1 cells were treated with T/S/Z or DMSO solvent control for 0, 24, 48, or 72 h. At the end of treatment, cell viability was determined by MTS assay. d Immunoblot analysis to detect phosphorylated RIPK3 (p-RIPK3), RIPK3, phosphorylated MLKL (p-MLKL), MLKL, and β-actin in lysates from CNE1 and CNE1-LMP1 cells treated with T/S/Z for 0, 24, 48, or 72 h. e TEM photomicrographs of CNE1 and CNE1-LMP1 cells treated with DMSO or T/S/Z. Black and white arrows denote plasma membrane rupture and cellular organelle swelling, respectively. Scale bars represent 5 μm. f CNE1 and CNE1-LMP1 cells were treated with T/S/Z, T/S/Z + Nec-1, T/S/Z + GSK’872, Nec-1, GSK’872, or DMSO solvent control for 72 h. Cells were then stained with Sytox Green and imaged by a fluorescent microscope. Scale bars represent 50 μm