Measurement of apoptosis and autophagy in PC-3 cells that received various treatments.

(A) Early apoptosis, detected using an Annexin V apoptosis detection kit, was measured using flow cytometry. Cells were treated with IR (4 Gy) and MP (25 μM) for 48 hrs. (B) Development of AVOs in PC-3 cells. Detection of green and red fluorescence in AO-stained cells using flow cytometry. (C) Quantification of AVOs with AO-stained cells treated with IR (4 Gy) or MP (25 μM) alone or in combination using flow cytometry. #, p<0.05, IR versus combined treatment. *, p<0.05, MP versus combined treatment. Data are presented as the mean ± standard deviation of three independent experiments. (D) EM microphotographs of PC-3 cells treated with IR (4 Gy) and MP (25 μM) for 48 hrs. The white arrows point to autophagic vacuoles and autolysosomes. (E) Western blotting for LC3-I, LC3-II, p62/SQSTM1 and Atg5–12 in PC-3 cells. Cells were treated with IR (4 Gy) and MP (25 μM) for 48 hrs.