Fig. 3
Apoptosis of cancerous 4T1 cells induced by Mn/N-HCN. A Intracellular ROS level. Scale bars = 50 μm. B AO staining. Fluorescence turns red once AO enters the cytoplasm due to compromised lysosomal integrity. Scale bars = 50 μm. C JC-1 analysis. At normal depolarized mitochondrial membrane potential. JC-1 molecules aggregate and emit red fluorescence. Scale bars = 50 μm. D Expression of apoptotic proteins revealed by immunoblotting, and (E) Statistic analysis normalized to actin expression. Note: different treatments: (I) Control, (II) H2O2 (100 μM), (III) Mn/N-HCN (50 μg/mL), (IV) Mn/N-HCN + H2O2, (V) Mn/N-HCN + H2O2 + 1064 nm laser (1 W/cm2). Statistical analysis was analyzed using Student's t-test for comparison. (*P < 0.05, **P < 0.01, ***P < 0.001, n.s. = not significant.)