Figure 2
Iso-GNA Induced Human NSCLC A549 Cell Death Was Apoptosis-Independent.
(A) and (B) A549 (5 × 105 cells per well) cells were seeded on 6-well plates for 24 h and then incubated with indicated concentrations of iso-GNA or cisplatin for 24 h. then (A) cells were stained with PI before cell cycle analysis by flow cytometry. (B) Cells were stained with Annexin V-FITC/PI before cell apoptosis analysis by flow cytometry. (C) Expression of caspase3 in A549 cells. Different concentrations of iso-GNA or cisplatin were incubated with A549 cells for 24 h, then western blot was performed to analyze caspase3 expression. (D) Caspase3 activity in A549 cells. Different concentrations of iso-GNA or cisplatin were incubated with A549 cells for 24 h and then subjected to analyze Caspase3 activity. (E) Caspase-independent cell death caused by iso-GNA. Different concentrations of zVAD.fmk was added to A549 cells for 6 h before the treatment of 10 μM iso-GNA or 30 μM cisplatin for 36 h, then cells were stained with trypan blue to analyze death ratio. Data of three independent tests were shown as means ± s.d. *p < 0.5; **p < 0.01, in comparison with the untreated group. The whole images of western blots are given in Supplementary Figure S6.
