Dose-dependent effects of MBCD on PIEZO1-GFP in stable cell line. (A) Pressure–response curves comparing the effects of different concentrations of MBCD on PIEZO1-GFP pressure sensitivity. Peak current values (I) are normalized to the maximal current (I_max) within the same recording and fitted with a Boltzmann distribution curve. (B) Midpoint activation pressures (P₅₀) producing 50% of the maximal current response from each Boltzmann curve analyzed in A (one-way ANOVA; **, P < 0.005; n.s., not significant; n = 3–9 cells). (C) Quantification of the mean steady-state current produced by each cell. The average current measured over the last 100 ms of the pressure stimulus was normalized to the maximal current within the same cell and expressed as a percentage. n = 3–7 cells per condition. The steady-state current for each cell is the average mean steady-state current from a minimum of three pressure pulses between −40 and −60 mmHg. (D) Latency time in milliseconds of the PIEZO1-GFP response measured between the start of negative-pressure pulse and the onset of the maximal current within the same recording (n = 3 cells, where each is the average of at least three consecutive pressure protocols). Average latencies are displayed for pulse pressures between −20 and −70 mmHg. All data represent mean ± SEM; statistical analysis between control and treatments was performed using one-way ANOVA; **, P < 0.005; ****, P < 0.0001.