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. 2018 Jul 16;9:720. doi: 10.3389/fphar.2018.00720

FIGURE 6.

FIGURE 6

GC-mediated generation of ROS causes apoptosis in leukemic cells. NAC pre-treated leukemic cells prevented GC-induced increase in sub-G0 fraction in K562 (A) and U937 (B) cells. K562 and U937 cells were pretreated with 10 mM NAC, subsequently treated with 25 μM GC as indicated for 24 h, and cell cycle fraction was measured by flow cytometry. The graph displays the mean ± SD of three independent of experiments. ∗P < 0.05 and ∗∗P < 0.001. NAC pre-treated leukemic cell prevented GC-induced apoptosis. K562 (C) and U937 (D) cells were pretreated with 10 mM NAC, subsequently treated with 25 μM GC as indicated for 24 h and apoptosis was measured by staining with fluorescein-conjugated annexin-V and propidium iodide (PI) and analyzed by flow cytometry. The graph displays the mean ± SD of three independent of experiments. ∗P < 0.05 and ∗∗P < 0.001. NAC pre-treated leukemic cells prevented GC-mediated loss of mitochondrial membrane potential. K562 (E) and U937 (F) cells were pretreated with 10 mM NAC, subsequently treated with 25 μM GC as indicated for 24 h and loss of mitochondrial membrane potential was measured by JC1 staining and flow cytometry. The graph displays the mean ± SD of three independent of experiments. ∗P < 0.05 and ∗∗P < 0.001. NAC pre-treated leukemic cells prevented GC-mediated activation of caspases. K562 (G) and U937 (H) cells were pretreated with 10 mM NAC, subsequently treated with 25 μM GC as indicated for 24 h and lysed cell extracts were separated on SDS–PAGE, transferred to PVDF membrane, and immunoblotted with an antibody against procaspase-3, cleaved caspase-3, PARP, and tubulin.