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

Journal: Investigative Ophthalmology & Visual Science

Article Title: MicroRNA 9 Is a Regulator of Endothelial to Mesenchymal Transition in Diabetic Retinopathy

doi: 10.1167/iovs.64.7.13

Figure Lengend Snippet: Primer Sequences Used for RT-qPCR

Article Snippet: Pecam1 (ab204527; Abcam, Cambridge, MA, USA) and Acta2 (NBP2-66429; Novus Biologicals, Littleton, CO, USA) ELISA kits were used according to the suppliers’ instructions.

Techniques: Sequencing

Journal: Investigative Ophthalmology & Visual Science

Article Title: MicroRNA 9 Is a Regulator of Endothelial to Mesenchymal Transition in Diabetic Retinopathy

doi: 10.1167/iovs.64.7.13

Figure Lengend Snippet: MiR-9 prevented diabetes-induced EndMT and diabetes-induced vascular leakage in mouse retinas. Streptozotocin-induced diabetes significantly reduced retinal mRNA expressions of endothelial markers ( A ) Pecam1 and ( B ) Cdh5 , and increased expressions of mesenchymal markers ( C ) Acta2 and ( D ) S100a4 in male WT mice. Such changes were prevented in diabetic M9 mice. Protein expressions of ( E ) Pecam1 and ( F ) Acta2 also followed the same pattern. ( G ) IgG staining was seen within the retinal capillaries of all mice ( arrow ). Diabetes caused leakage of IgG into the retinal layers of WT diabetic mice, resulting in intense (+++) staining throughout the retina when compared with the nondiabetic mice (+). Diabetic M9 mice showed minimal leakage of IgG into the retina and had staining intensity comparable to non-diabetic M9 mice (+). ND, nondiabetic; D, diabetic; M9, miR-9 transgenic (n = 6/group for mRNA, n = 3/group for protein, and n = 3/group for immunohistochemistry; molecular data normalized to WT-ND, mRNA data presented as ratio to β-actin mRNA, and protein data presented as ratio to total protein; * P < 0.05).

Article Snippet: Pecam1 (ab204527; Abcam, Cambridge, MA, USA) and Acta2 (NBP2-66429; Novus Biologicals, Littleton, CO, USA) ELISA kits were used according to the suppliers’ instructions.

Techniques: Staining, Transgenic Assay, Immunohistochemistry