Table 2.
Summary of average P50 values and slopes derived from Boltzmann fits of I/Imax versus pressure plots of all PIEZO1 variants and treatments performed in this study
P50 (−mmHg)Slope of Boltzmann α−1 (mmHg−1)∆G (kBT)
hPIEZO1-GFP 36.4 ± 3 14 ± 2.5 3.5 ± 0.4 
+ 0.1 mM MBCD 30.7 ± 7 13.3 ± 1.3 3.2 ± 0.5 
+ 1 mM MBCD 47.1 ± 0.6 8.3 ± 3.7 5.7 ± 1.3 
+ 5 mM MBCD 54.2 ± 1.1 13.3 ± 1 5.3 ± 1.2 
+ 100 µg/ml cholesterol 36.8 ± 2.5 14.4 ± 2 3.5 ± 0.2 
+ 80 µM dynasore 36.7 ± 4.8 16.3 ± 3.9 3.7 ± 1 
+ 25 µM DHA (24 h) 16 ± 2.8 12.3 ± 1.4 1.6 ± 0.4 
+ 25 µM DHA (48 h) 27 ± 1.8 10.3 ± 1.4 3.6 ± 0.8 
hPIEZO1 WT 39.6 ± 2.1 11.6 ± 2 4.4 ± 0.5 
+ 5 mM MBCD 42.2 ± 2.6 14.3 ± 2.1 3.7 ± 0.6 
hPIEZO1 R2456H 13.3 ± 7 15.1 ± 3 1.3 ± 0.6 
+ 5 mM MBCD 16.4 ± 5.6 14.1 ± 2.8 1.9 ± 0.5 
N2A mPIEZO1 17.8 ± 2.2 6.6 ± 1.6 4.7 ± 0.3 
+ 5 mM MBCD 22.5 ± 1.8 8.4 ± 1.7 4 ± 0.7 

Gating free energy calculations were estimated following Eq. 7 (∆G/kBT ≈ P50 ∙ α) in Hamill and Martinac (2001). Statistical difference between ∆G detected for PIEZO1-GFP versus 25 µM DHA (24 h), P = 0.0153, Student’s t test. All values are reported as mean ± SEM.

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