(A) Schematic depicting whole-cell recording setup with mechanical indentation stimulation and gravity perfusion.
(B) Indentation stimulus protocol (5 μm, top), voltage protocol (middle), and representative whole-cell current (bottom) from a HEK293t-P1Ko cell transiently transfected with wtP1 in with (black) and without (gray) 10 mM DTT in the bath.
(C and D) Magnification of indentation stimulus protocol and currents from (B) at −80 mV (C) and +80 mV (D).
(E) Structural model of Piezo1 cap highlighting cysteine pair A2328C and P2382C. Colors indicate three subunits of Piezo1.
(F) Indentation stimulus protocol (5 μm) and representative currents from a cell transfected with A2328C/P2382C at −80 mV (top) and +80 mV (bottom) with (black) and without (gray) 10 mM DTT in the bath.
(G) Mean peak current from 8–11 individual cells, normalized to the average of the first four peak currents in DTT, for wtP1 and A2328C/P2382C, at −80 mV (top) and +80 mV (bottom).
(H) Mean inactivation time constant (τ) from 8–11 individual cells for wtP1 and A2328C/P2382C at −80 mV (top) and +80 mV (bottom).
All data are mean ± SEM. See also Figures S2 and S3.