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. 2017 Nov 27;108(12):2462–2469. doi: 10.1111/cas.13421

Figure 4.

Figure 4

Apoptosis induction by pyruvate kinase isozyme M2 (PKM2) inhibitor (shikonin) and/or glutaminase (GLS) inhibitor (BPTES). (a) Representative examples of flow cytometric analysis. To clarify the induction of apoptosis, double staining of cells with annexin V (FITC) and propidium iodide (PI) was carried out. Cells staining annexin V‐positive and PI‐negative were considered to be apoptotic; cells staining annexin V‐negative and PI‐positive were considered to be viable. (b) Apoptosis rates of cancer cells by PKM2 inhibitor and/or GLS inhibitor. The apoptosis rates of all of hypoxia‐resistant cancer cell lines were significantly increased by shikonin treatment, whereas treatment with BPTES did not affect the apoptosis of cancer cells except NUGC‐3/hypo cells. Combined treatment with shikonin and BPTES significantly increased the apoptosis of cancer cell lines, except NUGC‐3 cells. Results are presented as means of four independent experiments; bars indicate SD. *P < 0.05, **P < 0.01, ***P < 0.001.