Fig. 2: Single-channel Properties of WT versus PBS Piezo1 Variants.
HEK293TΔP1 cell attached patch clamp experiments were performed without and with applied pressure (−30 mmHg) at –60 mV. a Single-channel current recordings of WT and PBS variant proteins S260R, S2211L and co-expressed S260R + S2211L, showing the closed (C) ~ 0 pA and open (O) ~−1.5 pA state of the channel before (gray) and after (red) pressure application (red arrow). The pipette solution contained 130 mM Na+ and 1 mM Ca++, the bath solution contained 140 mM K+ and 1 mM Mg++. b All-point current histograms of the single-channel recordings shown in (a) (red) acquired under –30 mmHg pressure. The area under the curve demonstrates the C and O states of the channel. X-axis is the current amplitude in picoamperes (pA). Y-axis is Normalized Counts (NC). c Steady state NPo of WT and PBS variants either without any applied pressure to the patch of membrane (white bars) or with −30 mmHg applied pressure (black bars). X-axis is pressure in mmHg. Y-axis is NPo. (WT n = 24, S260R n = 17, S2211L n = 12, S260R + S2211L n = 12). d Single channel conductance of WT and PBS variant channels obtained from panel B at –60 mV. Experimental replica numbers are shown in the bar graph (n > 12). e Steady state NPo at −30 mmHg where experimental replica numbers are shown in the bar graph (n > 12) and (f) mean open time of WT and PBS variants (n > 12). Statistical analysis was performed by using Student’s t test (c) and One-way ANOVA (d–f) where *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001 and ns = not significant. The data in bar graphs in (c–f) are presented as Mean ± SD.
