Extended Data Fig. 1. Intersectional marking and characterization of PROKR2^ADV neurons.

a, Schematic description of the intersectional genetic strategy for generating Prokr2^Adv-tdTomato mice, in which tdTomato expression was confined to PROKR2^ADV sensory neurons upon removal of two STOP cassettes from the ROSA26 allele by the Cre and Flpo recombinases. Advillin^Flpo drove Flpo expression restricted to sensory neurons, such that DRG neurons with developmental coexpression of PROKR2-Cre and ADVILLIN-Flpo were marked by tdTomato. b, Representative sections through suprarenal sympathetic ganglia, adrenal medulla where chromaffin cells were located, the colon that should contain enteric ganglia, and nodose ganglia from Prokr2^Adv-tdTomato mice. The intersectional genetic strategy only labeled one or two cells per section in nodose ganglia. Arrows indicate tdTomato⁺ fibers, but not somas, likely from DRG neurons or from those rare nodose sensory neurons. In other words, PROKR2^ADV-tdTomato did not label any autonomic neurons and adrenal chromaffin cells. n = 5 mice per group. c, L4-5 DRG sections from adult Prokr2^Adv-tdTomato mice. Top panels show double immunostaining of tdTomato (red) with markers for nonperptidergic neurons (IB4: isolectin B4), proprioceptors (PV: parvalbumin), TRKA-lineage neurons (TRKA), myelinated neurons (NEFH: the neurofilament heavy chain protein), and peptidergic neurons (CGRP: the calcitonin gene-related peptide). Bottom panels show triple staining of tdTomato with NEFH and CGRP. Arrows indicate co-localization. n = 5 mice for each marker. Data are shown as mean ± SEM. Scale bars, 100 μm.