A potential issue in our MD simulation was that we used the same number of lipids for the asymmetric bilayers to remove the effects of lipid numbers, which may lead to a somewhat frustrated state of the bilayer in the rectangular simulation box due to the size difference of the asymmetric lipids. To alleviate this concern, we implemented a correction by subtracting the lateral pressure of the system in the absence of calcium ions from the lateral pressure of each simulation scenario. This adjustment can effectively neutralize the effects of the potential frustration induced by the asymmetric lipid composition.