Fig. 2: PIEZO1 is functionally expressed in a subset of putative itch neurons.

a, Percentage of wild-type neurons responding to compounds (2,682 neurons from 3 mice). AITC, allyl isothiocyanate. b, Venn diagrams of response overlap from combined wild-type data in a and d, indicating the percentage of total neurons. c, Traces of representative calcium signals from b. Arrowheads indicate compound addition. d, Percentage of neurons responding in Piezo1^fl/fl;Pirt^Cre−/− (wild type; WT) versus Piezo1^fl/fl;Pirt^Cre+/− (KO) neurons (1,883 WT and 2,218 KO neurons from 2 mice per genotype). e, Area under the curve of Piezo1^+/+ versus Piezo1^GOF/GOF responses to 20 µM Yoda1 (Mann–Whitney: ****P < 0.0001, U = 102,979; n = 347 +/+ and 711 GOF/GOF neurons from 2 mice per genotype). f, Peak normalized F₃₄₀/F₃₈₀ ratio of Piezo1^+/+ versus Piezo1^GOF/GOF mice from data in e (Mann–Whitney: *P = 0.0142, U = 121,046; n = 347 +/+ and 768 GOF/GOF neurons from 2 mice). In e,f, the centre line denotes the median, the boxes are the 25th and 75th percentiles and the whiskers indicate 1.5 times the interquartile range. g, Representative immunohistochemistry (IHC) section (of three sections from two mice) of Ai9 ^fl/fl;Sst^Cre+/− DRG neurons showing native tdTomato (expressed in SST⁺ cells) with the indicated markers (scale bar, 100 µm). h, Dissociated Ai9 ^fl/fl;Sst^Cre+/− DRG neurons. i–k, Summary of mechanically activated (MA) current inactivation kinetics in whole-cell poke experiments after nucleofection of Ai9^fl/fl;Sst^Cre+/− DRG neurons with the indicated siRNA mix against non-targeting control siRNA or against Piezo1 (i; ****P < 0.0001, χ² = 23.92, degrees of freedom (df) = 3; n = 28 control and 32 Piezo1 siRNA cells), Piezo2 (j; *P = 0.0130, χ² = 10.78, df = 3; n = 32 control and 31 Piezo2 siRNA cells) and Piezo1 + Piezo2 (k; ****P < 0.0001, χ² = 36.21, df = 3; n = 30 control and 33 Piezo1 + Piezo2 siRNA cells). Chi-squared (χ²) tests were performed. IA, intermediately adapting; NR, non-responsive; RA, rapidly adapting; SA, slowly adapting. l–n, Representative 150-ms indentation traces (top) and MA currents (bottom) from i–j with nucleofection of indicated siRNA (from two mice each). All statistical tests are two-tailed where applicable. n indicates biological replicates (cells).

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