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We carried out transthoracic, two-dimensional, guided M-mode echocardiography in unanesthetized mice. Measurements were done using a Sequoia C256 (Siemens) with the 15 MHz linear-array transducer. Intact essence hemodynamic infinitesimal calculus was performed as described and relevant details are provided (Supplementary Methods online). These studies used a four-electrode pressure-volume catheter (model SPR-839, Millar Instruments) placed through the left ventricular apex in the open-chest, anesthetized shiner and positioned along the longitudinal axis to disk assembly publication by impedance and pressure sensation by micromanometry.