Research Paper Volume 15, Issue 11 pp 4794—4819

Figure 8. Induction of necroptotic cell death in PVT1-sh#/ PVT1-OE ESCA cells. (A) ECA-109 cells were treated with Nec-1 (50 μM) for 4 h and then treated with TSZ (30ng/ml TNF-α, 200nM Smac mimetics, 20μM zVAD). After 24 h of drug treatment, the morphological changes of treated cells were imaged under a phase-contrast microscope. (B) Western blot from ECA-109 cell was performed to detect RIP1, p-RIP1, MLKL, and p-MLKL protein levels. (C) KYSE-510 cells were treated with Nec-1 (50 μM) for 4 h and then treated with TSZ (30ng/ml TNF-α, 200nM Smac mimetics, 20μM zVAD). After 24 h of drug treatment, the morphological changes of treated cells were imaged under a phase-contrast microscope. (D) Western blot KYSE-510 cell was performed to detect RIP1, p-RIP1, MLKL, and p-MLKL protein levels.