Figure 2.

Cells were subjected to prolonged hypoxia (12 h) after early and late preconditioning (EPC and LPC), in the absence and presence of aprotinin (KLK1 selective inhibitor), and after exogenous bradykinin (Bk) administration (10⁻¹² and 10⁻¹¹ M), as described in the text. Apoptosis was assessed using Annexin V (green), and necrosis was assessed using Propidium Iodide (red) (PI) staining; nuclei were stained with DAPI (blue). The rates of apoptosis and necrosis were calculated by dividing the number of Annexin-V-positive/PI-negative cells and Annexin-V-positive/PI-positive cells, respectively, by the total number of nuclei detected with DAPI staining. The percentage of apoptotic and necrotic cell death was calculated from six independent experiments. Quantification of apoptosis and necrosis is reported in Table S1 of Supplementary Material. KLK1: tissue kallikrein. The size of the scale bar is 2000 µm.