mouse anti canine cd3 fitc  (Bio-Rad)

Journal: Scientific Reports

Article Title: Exploring the effect of canine cancer-associated fibroblasts on T cell dynamics through the CXCL12/CXCR4 axis modulated by TGF-β1

doi: 10.1038/s41598-025-16312-x

Figure Lengend Snippet: Detection of CXCR4 expression on canine T cells. ( A ) CXCR4 expression on canine CD3⁺CD8⁻ and CD3⁺CD8 + T cells were evaluated using flow cytometry analysis. Representative data from one healthy beagle and a cancer-bearing dog. Plots are gated on lymphocyte > single cells > Live cells > CD3 + cells > CD8 - or CD8 + cells . ( B ) CXCR4 expression on CD8 + T cells or CD8 - T cells in healthy dogs and dogs with cancer. Data are presented as median with IQR (Healthy, n = 10; Cancer, n = 26). * P < 0.05, *** P < 0.001.

Article Snippet: In addition, cells were stained with the following directly fluorochrome-conjugated anti-canine antibodies: FITC-conjugated mouse anti-dog CD3 (clone CA17.2A12; Bio-Rad Laboratories), PE-conjugated anti-CD4 antibody (clone YKIX302.9, Bio-Rad Laboratories) and APC-conjugated rat anti-dog CD8 (clone YCATE 55.9; Thermo Fisher Scientific).

Techniques: Expressing, Flow Cytometry

Journal: Scientific Reports

Article Title: Exploring the effect of canine cancer-associated fibroblasts on T cell dynamics through the CXCL12/CXCR4 axis modulated by TGF-β1

doi: 10.1038/s41598-025-16312-x

Figure Lengend Snippet: Effect of TGF-β1 on CXCR4 expression on T cells. ( A ) CXCR4 expression on CD3⁺CD4 + and CD3⁺CD8 + T cells after being cultured with the medium containing TGF-β1 (5 ng/mL) with or without the TGF-βRI inhibitor SB431542 (5 µM) for 5 days. Mean ± SD ( n = 3). Plots are gated on lymphocyte > single cells > Live cells > CD3 + cells > CD4 + or CD8 + cells . ( B ) CXCR4 expression on CD3⁺CD4 + and CD3⁺CD8 + T cells after being cultured with NF-CM or CAF-CM for 5 days. Mean ± SD (Control, n = 3; NF-CM, n = 5; CAF-CM, n = 8). * P < 0.05, *** P < 0.001, **** P < 0.0001.

Article Snippet: In addition, cells were stained with the following directly fluorochrome-conjugated anti-canine antibodies: FITC-conjugated mouse anti-dog CD3 (clone CA17.2A12; Bio-Rad Laboratories), PE-conjugated anti-CD4 antibody (clone YKIX302.9, Bio-Rad Laboratories) and APC-conjugated rat anti-dog CD8 (clone YCATE 55.9; Thermo Fisher Scientific).

Techniques: Expressing, Cell Culture, Control

Journal: Scientific Reports

Article Title: Exploring the effect of canine cancer-associated fibroblasts on T cell dynamics through the CXCL12/CXCR4 axis modulated by TGF-β1

doi: 10.1038/s41598-025-16312-x

Figure Lengend Snippet: CAFs attract CD8⁺ T cells through the CXCL12/CXCR4 axis. ( A ) Schematic of the Boyden-chamber migration assay. CXCR4⁺CD8⁺ T cells were seeded in the upper compartment; the lower compartment contained control medium, recombinant CXCL12 (100 ng/mL) or CAF-CM. Migrated cells were counted after 24 h. ( B ) Migration toward recombinant CXCL12, expressed as fold change versus control. Mean ± SD ( n = 3). ( C ) Migration toward CAF-CM, expressed as fold change versus control. Mean ± SD ( n = 3). Control, culture medium; AMD3100, 4 µM CXCR4 inhibitor. ( D ) Schematic of the adhesion assay using a cell culture insert in a 6 cm dish. CAFs and tumor cells (RCM-SO or NMTCC) were plated on opposite sides of the insert and cultured for 24 h; the insert was then removed and CXCR4⁺ T cells added for a further 24 h. ( E ) Density of T cells adhered per cm 2 to cancer cell lines versus CAFs after 24 h. Mean ± SD ( n = 4). RCM-SO, canine mammary gland carcinoma cell line; NMTCC, canine transitional cell carcinoma cell line. ( F ) IHC of serial sections from a canine pulmonary adenocarcinoma stained for CXCL12 (top) and CD3 (bottom). Dashed lines delineate stromal and tumor cell areas, and asterisks (*) indicate the stromal regions. Scale bars 50 µm ( G ) CD3⁺ T cell density in stromal and tumor regions of thyroid carcinoma (TC, n = 2) and pulmonary adenocarcinoma (LC, n = 4). Numbers after histo-type indicate CXCL12 expression score. Mean ± SD. ( H ) Ratio of stromal to tumor CD3⁺ T cell density in each case. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.

Article Snippet: In addition, cells were stained with the following directly fluorochrome-conjugated anti-canine antibodies: FITC-conjugated mouse anti-dog CD3 (clone CA17.2A12; Bio-Rad Laboratories), PE-conjugated anti-CD4 antibody (clone YKIX302.9, Bio-Rad Laboratories) and APC-conjugated rat anti-dog CD8 (clone YCATE 55.9; Thermo Fisher Scientific).

Techniques: Migration, Control, Recombinant, Cell Adhesion Assay, Cell Culture, Staining, Expressing