Fig 2: Mechanosensory neuron gene expression is altered in DRG neurons of Piezo2 cKO animalsA. UMAP plot of DRG neurons collected from 5 juvenile (P21-P24) control and 5 littermate Cdx2-Cre; Piezo2 cKO animals. Neurons are color-coded by computationally defined cluster identities. Molecular classes were manually assigned based on markers defined in the literature. See also Figure S5.B. UMAP plot of DRG neurons collected from juvenile (P21-P24) control (red) and Piezo2 cKO (blue) animals. Neurons are color-coded by genotype.C. UMAP plot of neurons assigned as Aδ-LTMRs (Cluster 17, see Supplemental Figure 5A), which were subsetted from the main dataset and re-analyzed. Control (red) and Piezo2 cKO (blue) neurons cluster partly by genotype.D. UMAP plot of neurons assigned as Aβ-LTMRs (Cluster 10), which were subsetted from the main dataset and re-analyzed. Control (red) and Piezo2 cKO (blue) neurons cluster partly by genotype.E. UMAP plot of neurons assigned as Mrgprd+ C-HTMRs (Clusters 0, 2, 8, 25), which were subsetted from the main dataset and re-analyzed. Control (red) and Piezo2 cKO (blue) neurons cluster partly by genotype.F. UMAP plot of neurons corresponding to Bmpr1b+ Aδ-HTMRs (Cluster 16), which were subsetted from the main dataset and re-analyzed. Control (red) and Piezo2 cKO (blue) neurons do not cluster by genotype.G. Number of differentially expressed (DE) genes between control and Cdx2-Cre; Piezo2 cKO DRG neurons of different molecularly defined classes. DE genes were identified using the Wilcoxon rank-sum test, adjusted p value <0.05, log-fold change ≥0.5.H. Top: Images of thoracic-level DRG sections from control or Cdx2-Cre; Piezo2 cKO animals stained with anti-CGRP (magenta) or IB4-conjugated (cyan) antibodies. Bottom: Images of thoracic-level DRG sections from control or Cdx2-Cre; Piezo2 cKO animals stained with anti-Kv4.3 (magenta) or anti-Th (cyan) antibodies. Scale bar = 100 μm.I. Images of lumbar-level DRG sections from control or Cdx2-Cre; Piezo2 cKO animals stained with RNAscope probes to detect Calb1 (magenta, a marker for Aβ-LTMRs) or Rem2 (cyan) transcripts. Scale bar = 50 μm.J. Quantification of the number of IB4+ cells in thoracic DRG sections for control or Cdx2-Cre; Piezo2 cKO animals. There was no difference between the two groups (unpaired t-test). Each data point indicates one animal; averages are shown by horizontal bars; error bars indicate the s.e.m.K. Quantification of the number of CGRP+ cells in thoracic DRG sections for control or Cdx2-Cre; Piezo2 cKO animals. There was no difference between the two groups (unpaired t-test). Each data point indicates one animal; averages are shown by horizontal bars; error bars indicate the s.e.m.L. Quantification of the number of TrkBGFP+ cells in thoracic DRG sections for control or Cdx2-Cre; Piezo2 cKO animals. There was a reduction in the mutants (**p<0.01, unpaired t-test). Each data point indicates one animal; averages are shown by horizontal bars; error bars indicate the s.e.m.M. Quantification of the number of Th+ cells in thoracic DRG sections for control or Cdx2-Cre; Piezo2 cKO animals. There was a reduction in the mutants (***p=0.0001, unpaired t-test). Each data point indicates one animal; averages are shown by horizontal bars; error bars indicate the s.e.m.N. Quantification of the number of Kv4.3+ cells in thoracic DRG sections for control or Cdx2-Cre; Piezo2 cKO animals. There was a reduction in the mutants (**p<0.01, unpaired t-test). Each data point indicates one animal; averages are shown by horizontal bars; error bars indicate the s.e.m.O. Quantification of the percentage of Calb1+ cells that are Rem2+ in lumbar DRG sections for control or Cdx2-Cre; Piezo2 cKO animals. There was a reduction in the mutants (*p<0.05, Welch’s t-test). Each data point indicates one animal; averages are shown by horizontal bars; error bars indicate the s.e.m.