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Figure 5 | Molecular Cancer

Figure 5

From: 6-Shogaol induces apoptosis in human leukemia cells through a process involving caspase-mediated cleavage of eIF2α

Figure 5

Analysis eIF2α associated ER stress pathway related proteins in the effect of 6S induced apoptosis in Jurkat cells. (a) Jurkat cells were treated with 15 μM of 6S for 0, 2, 4, 6, and 12 h or treated with various concentrations of 6S as indicated for 6 h. The total cellular extracts were subjected to western blot assay. β-actin was used as a loading control. Each blot is the representative result of three independent experiments. (b) Jurkat cells were pretreated with the caspase inhibitor Z-VAD-fmk (20 μM) for 1 h followed by treatment with 15 μM 6S for 12 h. Apoptosis was determined using flow cytometry. The graph shown represents the mean ± SD. for three separate experiments. The difference were significant at **p < 0.01. (c) Jurkat cells were treated with vehicle (DMSO 0.1%, v/v) alone or 6S (15 μM) or Z-VAD-fmk (20 μM) or the combination of 6S and Z-VAD-fmk for 12 h, western blot assay showed that Z-VAD-fmk markedly reduce 6S-induced apoptosis. Each blot is the representative result of three independent experiments. (d) Jurkat cells were treated with vehicle (DMSO 0.1%, v/v) alone or 6S (15 μM) or Sal (5 μM) or the combination of 6S and Sal for 12 h, cells were stained with Annexin V/PI, and the percentage of apoptotic cells was determined using flow cytometry. The graph shown represents mean ± SD for three separate experiments. The difference were significant at **p < 0.01. (e) Jurkat cells were treated with vehicle (DMSO 0.1%, v/v) alone or 6S (15 μM) or Sal (5 μM) or the combination of 6S and Sal for 12 h. Western blot assay showed that Sal potentiates 6S to induce Jurkat cells apoptosis through maintain the hyperphosphorylated state of eIF2α.

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