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ECG Recording ECGs were recorded continuously in deeply anesthetized rats for all three main leads, by positioning stainless steel electrodes on all four limbs using an ECG monitor with a 2090 programmer (Medtronic, Minneapolis, MN, USA) connected to a Waverunner LT342 digital oscilloscope (LeCroy, Chestnut Ridge, NY, USA) at 15 min, 24 h or 48 h ligation-time

While significant adverse events are uncommon at research-appropriate doses, it's prudent to monitor for any unexpected physiological changes