Journal: Cell reports

Article Title: CGRP signaling links tumor-associated pain to immune evasion in oral squamous cell carcinoma

doi: 10.1016/j.celrep.2026.116994

Figure Lengend Snippet: (A and B) Representative images of OSCC tumor serial sections stained with either anti-Tubb3 (top, green outline), anti-TRPV1 (middle, red outline), or anti-CGRP (bottom, blue outline) from a patient with (A) and without (B) CGRPα + nerve innervation. Positive stains are indicated by red (TRPV1) or blue (CGRP) arrows. Tumor margins are indicated by a gray dashed line. Image magnification: 6× (left) and 40× (right). Scale bar: 200 μm. (C) Quantification of the percentage of total TRPV1 and CGRP-IR nerve area relative to total Tubb3-IR nerve area across tumor tissue sections from 23 patients with HNSCC. Density is reported as a stacked bar graph. (D) Quantification of the percentage of total CD8 T cell density per square millimeter. (E) Representative images of OSCC tumor serial sections with either a large nerve bundle and low anti-CD8 or small nerve presence and high anti-CD8 immunoreactivity. Scale bar: 150 μm. (F and G) Simple linear regressions were run between patient-reported pain, percentage of total CGRP-IR nerve area relative to total Tubb3-IR, and CD8 + T cell density relative to tumor area. Pain was measured by the FACT-HN additional question 12, “I have pain in my mouth, throat or neck” (FACT-HN10). The response to this question is rated on a scale of 0 (not at all) to 4 (very much). Spearman correlation r coefficients are listed on each graph; p < 0.05. Patient demographics are located in .

Article Snippet: Slides were incubated overnight in PBS +/+ containing 1% bovine serum albumin and 0.1% Tween 20 and one of the following primary antibodies: Rabbit anti-CGRP (1:500, Cell Signaling Technologies), Rabbit anti-TRPV1 (1:500, Alomone Labs), Guinea pig anti-TRPV1 (1:500, Millipore).

Techniques: Staining

Journal: Cell reports

Article Title: CGRP signaling links tumor-associated pain to immune evasion in oral squamous cell carcinoma

doi: 10.1016/j.celrep.2026.116994

Figure Lengend Snippet: (A) Representative images of CGRP-IR in 300 μm optically cleared sagittal tongue sections from naive (left), MOC1 (middle), and MOC2 (right) tumor-bearing mice (10× magnification stitch, full focus z stack). Scale bar: 0.5 mm. (B) Representative images of co-staining with anti-CGRP and the pan-neuronal marker PGP9.5 in a 20 μm sagittal tongue section to demonstrate CGRP antibody specificity. Scale bar: 50 μm. (C) Representative images of neurite outgrowth in dissociated TG neurons in response to 24 h incubation in cell culture medium, MOC2 CM, or MOC2 medium + 1 μM anti-NGF monoclonal antibody (αNGF). A TG from one mouse was used for each treatment group, and the experiment was replicated five times ( n = 3 males, 2 females). For analysis, dissociated neurons were stained with anti-Tubb3 to visualize neurites and anti-NeuN to visualize cell bodies. Scale bar: 50 μm. (D–F) Integrative density of Tubb3 + neurites relative to the number of cell bodies in each field in response to medium and CM with or without αNGF; 12 areas of each chamber were imaged under 20× magnification, and analysis was completed within cell culture type (i.e., PEK, MOC1, and MOC2). Data are represented as mean ± SEM. One-way ANOVA; * p < 0.05, ** p < 0.01, *** p < 0.005, **** p < 0.001.

Article Snippet: Slides were incubated overnight in PBS +/+ containing 1% bovine serum albumin and 0.1% Tween 20 and one of the following primary antibodies: Rabbit anti-CGRP (1:500, Cell Signaling Technologies), Rabbit anti-TRPV1 (1:500, Alomone Labs), Guinea pig anti-TRPV1 (1:500, Millipore).

Techniques: Staining, Marker, Incubation, Cell Culture

Journal: Cell reports

Article Title: CGRP signaling links tumor-associated pain to immune evasion in oral squamous cell carcinoma

doi: 10.1016/j.celrep.2026.116994

Figure Lengend Snippet: (A) Representative image of a coronal tongue section from non-tumor bearing mice 1 week following treatment with vehicle (top) or RTX (bottom), stained with anti-CGRP to demonstrate loss of CGRP-expressing fibers following RTX treatment. Scale bar: 100 μm. (B) Schematic of the trigeminal CGRP release assay. (C) CGRP protein quantification in TG isolated from mice 1 week after RTX injection into the tongue ( n = 5 F/group). Data are represented as mean ± SEM. Independent t test, ** p < 0.01. (D) Representative ATF3 staining of in TRPV1-IR neurons in the trigeminal mandibular branch in TG sections from vehicle- and RTX-treated mice. Scale bar: 50 μm. (E) Percentage of ATF3 + TRPV1 + neurons relative to total TRPV1 + neurons in the V3 region from across 9 sections/mouse of mice treated with vehicle or RTX ( n = 3 vehicle, 4 RTX). Tissue was collected 7 days after treatment. There was no difference in the number of TRPV1 neurons between groups. Data are represented as mean ± SEM. Independent t test, *** p < 0.005. (F) RTX treatment did not affect body weight compared to vehicle-treated mice. Change in body weight was calculated as (post-treatment day 7) – (weight on first day of treatment). Independent t test. (G) Representative H&E-stained 5 μm tongue sections from vehicle- and RTX-treated mice 21 days after treatment. The tumor boundary is indicated by a black dashed line. Scale bar: 1 mm. (H) MOC1 tumor volume between groups over time ( n = 5 F/group). Data are represented as mean ± SEM. two-way ANOVA, * p < 0.05. (I and J) Example dot plots and quantification of CD3 + T cell subtype, CD19 + B cells, and NK1.1 + natural killer (NK) cells between groups ( n = 3 F/group). Within-subtype t test comparison, **** p < 0.0001.

Article Snippet: Slides were incubated overnight in PBS +/+ containing 1% bovine serum albumin and 0.1% Tween 20 and one of the following primary antibodies: Rabbit anti-CGRP (1:500, Cell Signaling Technologies), Rabbit anti-TRPV1 (1:500, Alomone Labs), Guinea pig anti-TRPV1 (1:500, Millipore).

Techniques: Staining, Expressing, Release Assay, Isolation, Injection, Comparison

Journal: Journal of Pain Research

Article Title: Electroacupuncture Alleviates KOA-Induced Pain and Cartilage Degeneration via NGF/TrkA Pathway

doi: 10.2147/JPR.S560506

Figure Lengend Snippet: EA intervention inhibits TrkA expression in CGRP + neurons in DRG of KOA model rats. ( A ) Representative immunofluorescence images of TrkA in DRG from saline group, MIA group, MIA + sham EA group and EA group. Scale bar = 200 µm. ( B ) Quantification of the number of TrkA + cells in DRG across the four groups (per group, n = 3). *p < 0.05; ns: not significant. ( C ) Representative images of double-labelling immunostaining of TrkA + with CGRP + , IB4 + , NF200 + in DRG sections from the saline, MIA, MIA + sham EA, and MIA + EA groups, scale bars = 200 µm. ( D) Pie chart summarizing the proportion of TrkA co-localization in IB4 + , CGRP + , and NF200 + neurons in KOA model rats. Three sections were analyzed per rat, and the results were averaged. ( E ) Quantification of the percentage of CGRP + neurons co-expressing TrkA in the DRG across the four groups (per group, n = 3). **p < 0.01, ***p < 0.001; ns: ns: not significant. ( F ) Quantification of the percentage of IB4 + neurons co-expressing TrkA in the DRG across the four groups (per group, n = 3). *p < 0.05; ns: not significant. ( G ) Quantification of the percentage of NF200 + neurons co-expressing TrkA in the DRG across the four groups (per group, n = 3).

Article Snippet: Goat anti-β-NGF antibody (3 μg/mL, R&D Systems, USA), goat anti-TrkA antibody (3 μg/mL, R&D Systems, USA), rabbit anti-PGP9.5 antibody (1:800, Abcam, UK), rabbit anti-TNF-α antibody (1:1000, Abcam, UK), rabbit anti-IL-1β antibody (1:1000, Abcam, UK), rabbit anti-CGRP antibody (1:400, Cell Signaling Technology, USA), mice anti-NF200 antibody (1:200, Cell Signaling Technology, USA), Isolectin B4 (BSI-B4, 1:1000, Sigma-Aldrich, USA).

Techniques: Expressing, Immunofluorescence, Saline, Immunostaining