Journal: Annals of Gastroenterological Surgery

Article Title: Intratumoral Fusobacterium nucleatum Drives Cancer‐Associated Fibroblasts Enrichment and Immune Exclusion in Esophageal Squamous Cell Carcinoma

doi: 10.1002/ags3.70116

Figure Lengend Snippet: Histopathological assessment of CAFs in ESCC. (a) Representative FISH images of ESCC tissues showing signals for F. nucleatum (red; FUS664), all bacteria (green; EUB338), and nuclei (blue; DAPI). Positive F. nucleatum signals were observed in cases that were identified as F. nucleatum –positive by qPCR. All images were acquired at ×200 magnification. (b) Representative FISH images of ESCC tissues obtained by laser scanning confocal microscopy, demonstrating intracellular signals of Fusobacterium nucleatum (red; FUS664) within tumor cells. Nuclei were counterstained with DAPI (blue). Images were acquired at ×400 magnification. (c) Representative FISH images of ESCC tissues showing signals for F. nucleatum (red; FUS664), all bacteria (green; EUB338), and nuclei (blue; DAPI). F. nucleatum signals were observed within tumor cells located in regions enriched with α‐SMA–positive CAFs. The boxed areas indicate regions of interest, imaged at ×200 magnification. ** p < 0.01.

Article Snippet: We used monoclonal mouse anti α‐SMA antibody (1:250; #56856; Cell Signal Technology) and RelA (1:800; #8242; Cell Signal Technology) as the primary antibody, following the manufacturer's protocols.

Techniques: Bacteria, Confocal Microscopy

Journal: Annals of Gastroenterological Surgery

Article Title: Intratumoral Fusobacterium nucleatum Drives Cancer‐Associated Fibroblasts Enrichment and Immune Exclusion in Esophageal Squamous Cell Carcinoma

doi: 10.1002/ags3.70116

Figure Lengend Snippet: Immunohistochemical analysis of NF‐κB activation and its association with F. nucleatum and CAFs in ESCC. (a) The proportion of NF‐κB–positive tumors was significantly higher in F. nucleatum –positive cases than in F. nucleatum –negative cases. (b) Representative immunohistochemical staining on adjacent serial sections of ESCC tissues showing stromal α‐SMA expression and nuclear RelA localization in tumor cells. These signals were observed in close proximity. Images were acquired at ×200 magnification. (c) Dual positivity for stromal α‐SMA and NF‐κB–positive in tumor cells was significantly enriched in F. nucleatum –positive tumors compared to F. nucleatum –negative tumors. (d) Summary of the results. F. nucleatum contributes to the progression of ESCC by inducing NF‐κB–mediated inflammatory signaling in tumor cells and promoting the activation of CAFs. ** p < 0.01.

Article Snippet: We used monoclonal mouse anti α‐SMA antibody (1:250; #56856; Cell Signal Technology) and RelA (1:800; #8242; Cell Signal Technology) as the primary antibody, following the manufacturer's protocols.

Techniques: Immunohistochemical staining, Activation Assay, Staining, Expressing