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. 2023 Jan 26;24(3):439–451. doi: 10.1038/s41590-022-01418-5

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© The Author(s) 2023

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 2 — a, Immunoblot showing the expression level of SNX25 in the lung of WT and Snx25^+/− mice. b, VF thresholds of WT and Snx25^+/− mice (WT, n = 19; Snx25^+/−, n = 33). P = 7.844 × 10⁻⁵. g, gram. c, Pain-related behavior time plotted for 5-min periods in WT (n = 6) and Snx25^+/− (n = 6) mice with injection of formalin into hind paws. s, second. d, Pain-related behavior time during phase 1 (0–10 min, P = 0.01) and phase 2 (20–60 min, P = 0.029) in mice as in c. s, second. e, VF thresholds plotted after SNI in WT and Snx25^+/− mice (WT, n = 4; Snx25^+/−, n = 7) at day 3 (P = 0.032), day 5 (P = 0.008) and day 7 (P = 0.04). g, gram. f, Representative immunoblots showing expression of TRPV1 and SNX25 in the DRG of WT and Snx25^+/− mice. g, Semi-quantitative analyses of immunoblots of TRPV1 and SNX25 in DRGs from WT (n = 5) and Snx25^+/− (n = 5) mice. TRPV1, P = 0.033; SNX25, P = 0.007. h, Confocal microscopy of the DRG stained with anti-TRPV1 antibody in WT and Snx25^+/− mice. Scale bar, 100 μm. Right, magnified views of boxed areas in the corresponding left panels. Representative of three independent experiments. Scale bar, 20 μm. i, Confocal microscopy of the DRG of WT and Snx25^+/− mice, stained with anti-TrkA antibody (left; scale bar, 100 μm) and quantification of mean TrkA fluorescence intensity (WT, n = 13; Snx25^+/−, n = 12 DRG sections from four different mice) (right). P = 0.042. Representative of three independent experiments. DAPI, 4,6-diamidino-2-phenylindole. j, Fluo-4 Ca²⁺ imaging of primary DRG neurons from an entire well dissociated from WT and Snx25^+/− mice (WT, n = 3; Snx25^+/−, n = 3). The arrow indicates the time when capsaicin was added to a well. P values are as follows: 448 s, P = 0.033; 469 s, P = 0.008; 490 s, P = 0.04; 511 s, P = 0.002; 532 s, P = 0.014; 553 s, P = 0.03; 574 s, P = 0.038; 595 s, P = 0.034; 700 s, P = 0.046; 742 s, P = 0.049; 805 s, P = 0.049. ex., excitation; em., emission. k, mRNA expression for pain-related factors in the DRG of WT and Snx25^+/− mice (WT, n = 3; Snx25^+/−, n = 3). Trpv1, P = 0.026; Scn9a, P = 0.032; Scn10a, P = 0.022. Results are represented as mean ± s.e.m. Statistical significance was calculated using two-tailed Student’s t-test (b–e, i and j) or two-tailed Welch’s t-test (g,k). *P < 0.05, **P < 0.01.

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