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. 2019 Jun 19;14(6):e0218659. doi: 10.1371/journal.pone.0218659

Fig 2. DR induces non-apoptotic cell death at low concentrations.

Fig 2

(A) DR was treated with various concentration for 24 h, and cell cycle analysis was performed using flow cytometry. (B) The cell cycle distribution was quantified and represented as a bar graph. (C) After 24 h incubation with the indicated concentrations of DR, the A549 cells were lysed and aliquots were assayed for in vitro caspase-8, - 9 and -3 activity. (D) After DR treatment for 24 h, the cells were stained with Annexin V. Early apoptotic Annexin V-positive cells were detected by flow cytometry. (E) After treatment of DR with the indicated concentrations, the cells were lysated and analyzed by western blotting. (F) Cells were co-treated with pan caspase inhibitor (Z-VAD-fmk, 20 μM) and cell viability were measured by MTT assay. Statistical differences were presented p<0.05 (*), p<0.01 (**), and p<0.001 (***) compared with the DR alone; p<0.01 (##) compared with the DMSO control.