Figure 4.
Evaluation of Piezo1 activity in BEL-A overexpressed wild-type and mutant cells using patch clamp. Raw traces of current amplitudes (A) and time course of the experiment (B) at 80 and −100 mV from an example overexpressed wild-type (WT) Piezo1 cell obtained before (precompound, red trace/symbols) and after (compound, blue trace/symbols) addition of 10 μM Yoda1, using a voltage ramp protocol (−100 to 80 mV, 400 ms, holding potential −30 mV) and inhibited by 30 μM GdCl3 (block, green trace/symbols). (C) Evaluation of current increase on Yoda1 addition (Yoda1 induced current, IYoda1− IES, pA/pF) of responding/nonresponding cells identified from BEL-A overexpressed wild-type and mutant Piezo1 constructs (n = 11/77 overexpressed WT; n = 21/84 Glu2392Lys; n = 13/80 Gly2394Ser; n = 15/89 Glu2407Gln; n = 15/80 Glu2407Lys; n = 19/85 Arg2245Gln). (D) Evaluation of current increase on Yoda1 addition in the presence of TRAM-34 (Yoda1 + TRAM-34 induced current, IYoda1− IES, pA/pF) of responding cells identified from BEL-A overexpressed wild-type and mutant Piezo1 constructs (n = 48/82 overexpressed WT; n = 54/86 Glu2392Lys; n = 62/87 Gly2394Ser; n = 47/93 Glu2407Gln; n = 47/77 Glu2407Lys; n = 45/84 Arg2245Gln).

Evaluation of Piezo1 activity in BEL-A overexpressed wild-type and mutant cells using patch clamp. Raw traces of current amplitudes (A) and time course of the experiment (B) at 80 and −100 mV from an example overexpressed wild-type (WT) Piezo1 cell obtained before (precompound, red trace/symbols) and after (compound, blue trace/symbols) addition of 10 μM Yoda1, using a voltage ramp protocol (−100 to 80 mV, 400 ms, holding potential −30 mV) and inhibited by 30 μM GdCl3 (block, green trace/symbols). (C) Evaluation of current increase on Yoda1 addition (Yoda1 induced current, IYoda1− IES, pA/pF) of responding/nonresponding cells identified from BEL-A overexpressed wild-type and mutant Piezo1 constructs (n = 11/77 overexpressed WT; n = 21/84 Glu2392Lys; n = 13/80 Gly2394Ser; n = 15/89 Glu2407Gln; n = 15/80 Glu2407Lys; n = 19/85 Arg2245Gln). (D) Evaluation of current increase on Yoda1 addition in the presence of TRAM-34 (Yoda1 + TRAM-34 induced current, IYoda1− IES, pA/pF) of responding cells identified from BEL-A overexpressed wild-type and mutant Piezo1 constructs (n = 48/82 overexpressed WT; n = 54/86 Glu2392Lys; n = 62/87 Gly2394Ser; n = 47/93 Glu2407Gln; n = 47/77 Glu2407Lys; n = 45/84 Arg2245Gln).

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