Fig. 5. TSP-1 inhibits PGE2-mediated nociceptor sensitization.

a, t-SNE plot of normalized Thbs1 expression in immune cells from healthy (CL) and inflamed skin following zymosan, incision and UV burn injury at T_max. b, Dot plot showing the expression of TSP-1 receptors (Cd47, Cacna2d1, Lrp1, Sdc1, Itga6, Itga4, Itgav and Cd36) in DRG neuron subtypes (cLTMR1, NF1, NF2, NF3, NP, p_cLTMR2, PEP1 and PEP2) in wild-type (WT) healthy mouse lumbar DRG⁴. c, The expression of CD47 on frozen wild-type or Cd47^−/− DRG neuron sections stained with PGP9.5. Scale bar, 100 μm. Representative of two independent experiments. d, Fura-2-based calcium imaging in cultured sensory neurons from DRG obtained from WT male mice treated with SES, 1 µM PGE2 or 1 µM PGE2 + 200 ng ml⁻¹ TSP-1 for 7 min, followed immediately by 100 nM capsaicin, 100 nM capsaicin + 1 µM PGE2 or 100 nM capsaicin + 1 µM PGE2 + 200 ng ml⁻¹ TSP-1, respectively for 30 s, a 5 min SES wash and treatment with 1 µM capsaicin for 30 s. SES was used as the recording solution. Treatments were applied during live imaging using a gravity-based perfusion system. Frames were captured every 3 s. Intensity traces of ratio of 340/380 are plotted. e, dF/F calculated as (F₁ − F₀)/F_0, where F₁ is the peak response within 40 s of treatment with 100 nM capsaicin and F₀ is the average of 10 s before the PGE2-treatment time point. SES, n = 80; PGE2, n = 55; PGE2 + TSP, n = 42. All TRPV1⁺ neurons are from one experiment consisting of three separate recordings for each condition. Data are represented as mean value ± s.e.m. Unpaired two-tailed t-test was performed. P < 0.05 was considered significant. An individual dot represents a neuron that was included in the analysis. Data are representative of three independent experiments.