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. Author manuscript; available in PMC: 2023 May 13.
Published in final edited form as: Circ Res. 2022 Apr 6;130(10):1531–1546. doi: 10.1161/CIRCRESAHA.122.320827

Fig. 5: Genetic deletion and RR abolish Piezo1 activity.

Fig. 5:

(a, b) Representative traces from a cEC from an EC-specific Piezo1-knockout (Piezo1EC-KO) mouse (a) and a cEC from a control mouse (b). Recordings were obtained in the cell-attached mode at a holding potential of −50 mV before (0 mmHg) and after the application of negative pressure (−10 mmHg) onto the patch. (c) Scatter plots of the open probability (NPO) of pressure-induced Piezo1-like single-channel openings in Piezo1EC-KO and control cECs (**P < 0.01, Wilcoxon test; Piezo1EC-KO, n = 6 cECs/4 mice; Control, n = 9 cECs/3 mice). (d) Scatter plots of the open probability of Yoda1-induced channel openings in Piezo1EC-KO and control cECs (**P < 0.01, Mann-Whitney test; Piezo1EC-KO, n = 12 cECs/4 mice; Control, n = 12 cECs/4 mice). ns indicates not significant. Horizontal black lines depict means and error bars are SEM. (e) Representative traces of cell-attached, Yoda1-induced, single-channel inward currents in the presence of 1, 10 or 30 μM RR, recorded at a holding potential of −50 mV. Right insets: Magnified channel openings of the marked segments. Traces are representative of 7 cECs (1 μM RR), 6 cECs (10 μM), and 4 cECs (30 μM). (f) Open-time histograms in the presence of 1 or 5 μM RR in the pipette solution. Lines show a single exponential fit to a Marquardt algorithm, yielding mean open times of 8.9 ms (1 μM) and 3.3 ms (5 μM). (g) Concentration-dependent decrease in open time and estimated IC50 (n = 4, 10, 12, 3, 1 cECs at 0, 1, 5, 10, 30 μM RR, respectively). (h) Reciprocal of mean open time (1/ƬO) plotted against the external [RR].