(A) Representative fluorescent images of processed contralateral (vehicle-treated) and ipsilateral (CFA-treated) L5 DRG. Shown are RNAscope stainings with specific probes for TRPM3 (left), TRPA1 (middle) or TRPV1 (right) (green). Retrogradely labeled neurons were identified based on the WGA-AF647 staining (magenta), and the blue color represents the nuclear marker DAPI. Retrogradely labeled neurons in the boxed areas are shown at double magnification, along with RNAscope staining for the neuronal marker Pgp9.5 (yellow), which was used to delineate neuronal cell bodies. (B,D,F) Quantification of the number of RNAscope dots per DRG neuron for TRPM3, TRPA1 and TRPV1, comparing retrogradely labeled (red) and unlabeled (black) sensory neurons, from the contralateral and ipsilateral L5 DRG. Values are presented as mean along with the 95% confidence interval. The data of the individual cells are shown in Figure 1—figure supplement 2. Statistical comparisons between groups were made using Kruskal-Wallis ANOVA with Dunn’s posthoc test. (C,E,G) Fraction of DRG neurons that showed a positive RNAscope signal (≥5 dots) for the three tested channels. Values are presented as mean ± SEM along with data points from the individual mice. Statistical comparisons between groups were made using one-way ANOVA with Holm–Šidák post-hoc test. Data are from six mice. The total numbers of analyzed neurons were for TRPM3: 752 ipsilateral and 1299 contralateral; for TRPA1: 954 ipsilateral and 947 contralateral; for TRPV1: 1054 ipsilateral and 995 contralateral.