Protocol for the detection of mechanosensitive response of proteins in cultured cells under compressive stress

Here, we present a protocol to detect mechanosensitive responses of proteins in cells under compressive stress. We describe steps for preparing elastic gels to compress cells grown on an imaging chamber. We then detail procedures for imaging proteins at the cell cortex using high-resolution confocal microscopy. The protocol can be applied to examine the mechanosensitive response of fluorescently tagged proteins in mitotic cells or round interphase cells adhering to the imaging surface. For complete details on the use and execution of this protocol, please refer to Wang et al.¹.