Extended Data Fig. 5: Basic spinal respiratory parameters and lactate abundance in the absence of PTG (related to Fig. 5).

(a-c) Seahorse mitochondrial-stress test in spinal cord dorsal horn slices comparing Oxygen Consumption Rate OCR (a), Extracellular Acidification Rate ECAR (b) and mitochondrial parameters (c) between WT (N = 37 slices/5 mice) and gPTG^−/− (N = 40/5). (d) Representative images of the neuronally-expressed calcium indicator jRGECO1a.NLS.Flag showing fluorescence prior to (baseline) and during (HiK) stimulation with high-potassium (25 mM) aCSF. (e) Representative trace showing the percent change in fluorescence intensity over the course of the imaging period within a large region of interest encompassing all visibly labeled cells within the dorsal horn (left) and peak amplitudes of nuclear calcium responses measured within 2 minutes after the start of stimulation in 29 slices from N = 4 animals (right). (f) Effect of 10 μM capsaicin and 25 mM KCl on ECAR in WT spinal cord dorsal horn slices. Data normalized to the average baseline of each condition (N = 7/1). (g) Change of ECAR produced by the stimulus (capsaicin or KCl) measured as change of area under the curve (AUC) derived from data shown in (f; One-way ANOVA N = 7/1). (h) Lactate content after formalin pain stimulus, measured by mass spectrometry and normalized to naïve animals. One-way ANOVA with Tukey’s post hoc test (N = 5). (i) Lactate content in spinal cord dorsal horn tissue of WT and gPTG^−/− mice before and at indicated time points after CFA injection. Two-way ANOVA (N = 4). Data represent mean ± s.e.m.