The effects of mild and aggressive mechanical stimulations on LRP6, JNK signaling, eNOS production, and calcium ions in cardiomyocytes. (A,B) Piezo1 distribution observed to be different after subjected to mild and aggressive mechanical stimulations. (C) Cardiomyocytes subjected to 5% stretching for 24 h increased the LRP6/β-catenin (Wnt pathway) signaling. The SERCA2 is known to act as calcium transporter on the sarco/endoplasmic reticulum to promote cell contractility. In this study, SERCA was detected in the cardiomyocytes. (D) Piezo1 inhibition reduced calcium release at 5%. (E) At 25%, calcium ion release and T-eNOS protein level were reduced compared to 5%. (F) Piezo1 inhibition at 25% reduced calcium ion release; however, significantly increased LRP6/β-catenin, and JNK.