Deletion of Ca_V1.2 in NaV1.8⁺ nociceptors reduces PKA-II activity. (A) Expression pattern of Na_V1.8 (Scn10a), Ca_V1.2 (Cacna1c), and RIIβ (Prkar2b) in subgroups of DRG neuron determined by single-cell RNA-seq (Zeisel et al., 2018). (B) Conditional mouse model to delete Ca_V1.2 in Na_V1.8-expressing DRG neurons. (C and D) Distribution of UCHL1 and RIIβ expression levels in DRG neurons of cKO mice and respective controls lacking Cre recombinase (Ctrl). (E) Total numbers of viable DRG neurons after overnight culture determined by HCS microscopy (n = 3 females and 2 males per genotype). (F) Single-cell data and mean intensities obtained using a Ca_V1.2-specific antibody (clone N263/31) indicating down-regulation of Ca_V1.2 (n = 10 cultures from three mice per genotype, >6,000 neurons per genotype, Student’s t test). The primary antibody was omitted in respective controls (w/o AB). (G) Basal genotype difference of pRII intensity in all solvent stimulated RIIβ⁺ control neurons (n = 3 females and 2 males per genotype, >15,000 neurons/condition, Student’s t test). (H) Dose-dependent induction of pRII intensity by KCl (0–80 mM) in cKO and Ctrl mice (n = 3 females and 2 males per genotype; genotype effect: F_4,62 = 5.6, extra-sum-of-squares F test). (I) Dose-dependent induction of pRII intensity by 10 mM KCl after 10 min preincubation with (S)-(-)-Bay K 8644 (0–5 µM) in cKO and Ctrl mice. (n = 3 females and 2 males per genotype; genotype effect: F_4,62 = 13.9, extra-sum-of-squares F test). (J) Single-cell data of selected condition shown in I. Values in G–I are means ± SEM; *, P < 0.05; ***, P < 0.001.