Fig. 4.

Cholesterol effect on Piezo1 clustering and channel kinetics. (A) Super-resolution microscopy shows the channels clustering together at the nanoscale (top). These clusters are dependent on the cholesterol content of the membrane (bottom). (B) Leaching cholesterol from the membrane changes sensitivity and gating kinetics. Under pressure application in a membrane patch (right control) the current decays as the channel inactivates. This behavior is lost if cholesterol is removed (top). Boltzmann distribution function showing dependence of Piezo1 currents on negative pressure (suction) applied to the cell-attached membrane patch of HEK293 cells. Piezo1 channels in cells treated with 5 mM MBCD exhibit reduced mechanosensitivity as indicated by a shallowed slope of the Boltzmann function (bottom left). The half activation pressure of Piezo1 channels increased significantly in membrane patches treated with MBCD (bottom right) (modified from Ridone et al. (2019))