Journal: Neurobiology of Pain

Article Title: Peripheral immune response and axonal degeneration in the hind paw skin of mice with experimental autoimmune encephalomyelitis

doi: 10.1016/j.ynpai.2025.100207

Figure Lengend Snippet: Temporal dynamics of GFRα2 fiber innervation. (A-F) Representative images of PGP9.5 staining in the dermal-epidermal junction in male (A-C) and female (D-F) mice at various disease time points. (G) Quantification of GFRα2+ fiber crossings per mm of dermal–epidermal length. **** p < 0.0001, Two-way ANOVA, Tukey’s multiple comparisons test.

Article Snippet: Primary antibodies included: CD45 (BD Biosciences 550539, 1:500), CD11b (Abcam ab133357, 1:200), CD11c (Abcam ab219799, 1:200), Ly6B.2 (Abcam ab53457, 1:200), CD4 (Abcam ab183685, 1:500), CD8 (Abcam ab217344, 1:500), PGP9.5 (EnCor Biotechnology 187–010116, 1:500), CGRP (Millipore Sigma PC205L-100UL, 1:500), and GFRα2 (R&D Systems AF429, 1:200).

Techniques: Staining

Journal: Nature

Article Title: Maternal stress triggers early-life eczema through fetal mast cell programming

doi: 10.1038/s41586-025-09419-8

Figure Lengend Snippet: a , Cumulative time spent not responding to the tape (no-response time, seconds). b,c , Averaged time courses ± SEM of tape-directed bouts (b) and AUC (c) of CT (n = 8 M/7 F) and PS (n = 9 M/6 F) offspring. d , Ex vivo DRG neuron stimulation and calcium imaging. Percentage (%) of responsive neurons. e-g , Traces of the calcium-induced changes of fluorescence exclusively in responding neurons after addition of capsaicine (e, n = 35 [CT], 21 [PS]), b-alanine (f, n = 97 [CT], 57 [PS]) and MRS2365 (g, n = 87 [CT], 131 [PS]), and amplitude of the neuronal response measured as AUC of the highlighted duration (for 20 s after the addition of the stimulus), normalized per cell (upper panel) or per experiment (lower panel). Each triangle is a responding neuron, and each dot represents an experiment including 2 mice per group (d-g). h-j , Representative confocal microscopy images of neurofilament H (NFH, h) and GDNF Family Receptor Alpha 2 (Gfrα2, i) in glabrous skin sections and filament length (µm, j) per mm 2 in steady state W8 CT (n = 6) and PS (n = 7) offspring. Scale bars, 30 µm (cropped to 7 µm). k,l , Representative confocal microscopy images of Tubβ3, TH, IB4 and Substance P staining (k) and associated percentages (l) among all Tdt + neurons, in DRG from Nav1.8-Cre + ;Tdt CT (n = 6) and PS (n = 6 [TH + ], 7 [IB4 + , SP + ]) at steady state. Scale bars, 200 µm (cropped to 30 µm). Data are from at least two or three independent experiments. Bars represent mean values, and each dot or square corresponds to a single mouse (a,j,l). Mean + SEM; *P < 0.05, **P < 0.01, ***P < 0.001, ns: not significant; Unpaired t-test (a, c), Mann-Whitney test (d-g, j, l), two-sided. Illustrations in d were created using BioRender ( https://biorender.com ).

Article Snippet: Frozen or paraffin-embedded mouse skin sections pretreated using a heat-induced epitope retrieval method (in 10 mM sodium citrate buffer (pH 6.0)) were permeabilized for 30 min in PBS 0.5% (w/v)% BSA and 0.3% Triton X-100 and incubated overnight at 4 °C with fluorophore-coupled antibodies or unconjugated antibodies: anti-Filaggrin (Ozyme; BLE905801; 1:50), anti-Loricrin (Ozyme; BLE905101; 1:50), anti-mouse Keratin 6A (Ozyme; BLE905701; 1:50), Anti-Claudin 1 antibody (Abcam; ab15098; 1:50), Alexa Fluor 647 anti-mouse CD45 (BioLegend; 103124; 1:400), anti-TH (Sigma; AB1542; 1:100), Alexa Fluor 647 anti-Tubulin β3 (BioLegend; 801210; 1:50), anti-human/mouse GFRα2 (R&D Systems; AF429; 1:200), anti-mouse NFH (Sigma; AB1989; 1:1,000), anti-GFP (Proteintech; pabg1; 1:200), anti-RFP (Takara; 632496; 1:200), Anti-Substance P (Merck Millipore; MAB356; 1:200) and anti-αSMA (Thermo Fisher Scientific; 14-9760-82; Clone 1A4; 1/100).

Techniques: Ex Vivo, Imaging, Fluorescence, Confocal Microscopy, Staining, MANN-WHITNEY