Extended Data Fig. 10: Breathing changes in cell-specific PIEZO knockout mice.
a, Representative images of Piezo2 RNA in situ hybridization (magenta) in NEBs and vagal ganglia, NEBs visualized by NCAM immunochemistry (cyan), scale bar: 50 μm. b, Number of total NEB cells (NCAM1 immunoreactivity) and Piezo2-expressing NEB cells from mice in a, mean ± sem, dots: individual sections, n = 16 sections from 2 Control mice, 15 sections from 3 NEB-ΔPIEZO2 mice, one-way ANOVA with Bonferroni post hoc test: Piezo2 NEB-ΔPIEZO2 vs. all comparisons, p < 0.0001; all other comparisons, ns. c, Quantification of gasps following nebulized methacholine delivery in urethane-anesthetized mice, mean ± sem, dots: individual animals, n = 9 Control, 11 NEB-ΔPIEZO2, 3 Vagal-ΔPIEZO1/2, one-way ANOVA with Tukey post hoc test: F(2, 20) = 5.923, p = 0.0095; pairwise comparisons: Control versus NEB-ΔPIEZO2, p = 0.0198; Vagal-ΔPIEZO1/2 versus NEB-ΔPIEZO2, p = 0.0457; all other pairwise comparisons, ns. d, Quantification of gasps by whole body plethysmography during normoxia (21% O2), hypoxia (12% O2), and hypercapnia (5% CO2) (7 min), mean ± sem, dots: individual animals, n = 5 control, 9 NEB-ΔPIEZO2, two-way ANOVA with Bonferroni post hoc test: Condition x Genotype F(2, 24) = 0.02893, p = 0.9715; Control versus NEB-ΔPIEZO2, all pairwise comparisons, ns. e, Quantification (right) of breathing measurements made by whole body plethysmography (cartoon, left), mean ± sem, dots: individual animals, n = 14 Control, 11 NEB-ΔPIEZO2, 6 Vagal-ΔPIEZO1/2, one-way ANOVA with Bonferroni post hoc test: Gasps, F(2, 28) = 9.9558, p = 0.0007, Control versus NEB-ΔPIEZO2, p = 0.0104, Vagal-ΔPIEZO1/2 versus NEB-ΔPIEZO2, p = 0.0010; Tidal Volume, F(2, 28) = 7.263, p = 0.0029, Vagal-ΔPIEZO1/2 versus NEB-ΔPIEZO2, p = 0.002; all other ANOVA results and pairwise comparisons, ns. f, Quantification of respiratory mechanics performed in NEB-ΔPIEZO2 (top, orange, n = 6), Vagal-ΔPIEZO1/2 (bottom, blue, n = 3) mice and control littermates without Cre expression (grey, n = 4 left, 5 right), mean ± sem, dots: individual animals, unpaired t test, NEB-ΔPIEZO2 left-to-right: p = 0.0384, p = 0.0314, p = 0.0285, p = 0.0223; Vagal-ΔPIEZO1/2: p = 0.0040, p = 0.0106, p = 0.0022, p = 0.0010.
