Experimental protocol.
ischemia
ISCHEMIA
After the start of perfusion, all hearts were allowed to stabilize for at least 30 minutes and baseline LVDP was registered. The duration of global ischemia, induced by clamping the inflow tubing, was 40 minutes for mouse hearts and 25 minutes for rat hearts (Additional experiments were performed using pharmacological stimulation of sGC in RBC from WT and sGC-KO mice. In these experiments, the RBCs were incubated with the sGC stimulator BAY 41-2272 (10 μM; Sigma-Aldrich), the PDE5 inhibitor sildenafil (100 μM; Sigma-Aldrich) and the NO donor DEA/NO (200 μM; Sigma-Aldrich) for 20 minutes before being administered to the isolated heart. Isolated RBCs from WT mice were also incubated with the sGC stimulator vericiguat (10 μM Bayer AG) for 1 hour. After centrifugation (1,000