Journal: Cancer Science

Article Title: Identification of intravascular tumor microenvironment features predicting the recurrence of pathological stage I lung adenocarcinoma

doi: 10.1111/cas.12219

Figure Lengend Snippet: Five‐year rates of recurrence in patients with stage I adenocarcinoma according to the immunohistochemical staining scores of intravascular tumors

Article Snippet: E‐cadherin (NCH‐384; Dako Cytomation, Glostrup, Denmark), CD44 (DF1485; Novocastra, Newcastle, UK), CD44 variant 6 (VFF‐7; Acris Antibodies, Herford, Germany), and vimentin (VIM 3B4; Progen Biotechnik, Heidelberg, Germany) were used as EMT‐related markers.

Techniques: Immunohistochemical staining, Staining, Variant Assay

Journal: Cancer Science

Article Title: Identification of intravascular tumor microenvironment features predicting the recurrence of pathological stage I lung adenocarcinoma

doi: 10.1111/cas.12219

Figure Lengend Snippet: Immunohistochemical staining of tumor tissue in patients with lung adenocarcinoma with intratumoral vascular invasion. Stars indicate elastic fibers with positive Victoria blue van Gieson staining. (a,b) Intravascular stromal cells stained with (a) H&E and (b) immunohistochemical staining for vimentin. The arrows point to round or spindle‐shaped, brown‐stained, non‐cancerous cells. The staining results were negative for cancer cells. (c,d) E‐cadherin expression in cancer cells. (e,f) CD44 expression in cancer cells. (g,h) CD44 variant 6 expression in cancer cells. (i,j) Vimentin expression in cancer cells.

Article Snippet: E‐cadherin (NCH‐384; Dako Cytomation, Glostrup, Denmark), CD44 (DF1485; Novocastra, Newcastle, UK), CD44 variant 6 (VFF‐7; Acris Antibodies, Herford, Germany), and vimentin (VIM 3B4; Progen Biotechnik, Heidelberg, Germany) were used as EMT‐related markers.

Techniques: Immunohistochemical staining, Staining, Expressing, Variant Assay

Journal: bioRxiv

Article Title: Identification of SPP1-positive macrophages by single-cell spatial analysis in human lung tissues with mycobacterial infection

doi: 10.1101/2024.09.12.612778

Figure Lengend Snippet: Coexpression and ligand-receptor assay in InSituCor. A. Heatmap of coexpressed transcripts across different cell types. Coexpression of FN1, SPP1, and MARCO was specifically observed in macrophage_d (highlighted by the broken red line). B. Conditional correlation of ligand-receptor pairs in the ligand-receptor assay. The SPP1-CD44 and FN1-CD44 pairs show strong conditional correlations. C. Correlation network of SPP1 and CD44-associated transcripts. D. Spatial images showing SPP1-CD44 correlations in MTB and NTM tissues. Yellow indicates high correlation in the images (left: MTB1, right: NTM2). E. Spatial images from CosMx™ analysis of MTB and NTM samples. High expression of SPP1, CD44, and both are indicated by yellow, blue, and red dots, respectively.

Article Snippet: Anti-SPP1 rabbit polyclonal antibody (HPA027541, Sigma-Aldrich, St. Louis, MO) and anti-CD44 mouse monoclonal antibody (UMAB134, OriGene Technologies, Rockville, MD) were used as primary antibodies.

Techniques: Expressing

Journal: bioRxiv

Article Title: Identification of SPP1-positive macrophages by single-cell spatial analysis in human lung tissues with mycobacterial infection

doi: 10.1101/2024.09.12.612778

Figure Lengend Snippet: Expression of SPP1 and CD44 in macrophages around granuloma tissue with mycobacterial infection. A. Dot plot showing the expression of SPP1, MARCO, FN1, and CD44 across all cell types. B. Immunohistochemistry for SPP1 and CD44 in tissues from patients with MTB (upper panels) and NTM (lower panels) infections. C. Immunofluorescence and CosMx™ spatial images for SPP1 and CD44. SPP1 was expressed in the cytoplasm of multinuclear giant cells (immunofluorescence assay, left panels). CD44 expression was observed on the membrane of multinucleated giant cells and their neighboring cells. CosMx™ spatial images (right panels) showed that macrophage_d expresses both SPP1 and CD44. Colors representing different cell types are shown in the bottom right panel.

Article Snippet: Anti-SPP1 rabbit polyclonal antibody (HPA027541, Sigma-Aldrich, St. Louis, MO) and anti-CD44 mouse monoclonal antibody (UMAB134, OriGene Technologies, Rockville, MD) were used as primary antibodies.

Techniques: Expressing, Infection, Immunohistochemistry, Immunofluorescence, Membrane

Journal: bioRxiv

Article Title: Identification of SPP1-positive macrophages by single-cell spatial analysis in human lung tissues with mycobacterial infection

doi: 10.1101/2024.09.12.612778

Figure Lengend Snippet: Analysis of cellular communication among cell types using CellChat. A and B. Circular plots showing interactions among cell types in all pathways (A) and specifically in the SPP1 signaling pathway (B) across all seven cases analyzed with CosMx™. C. Circular plots depicting interactions among cell types in the SPP1 signaling pathway in each individual case. D. Histogram showing the relative contribution of each ligand-receptor (L-R) pair across all seven cases analyzed with CosMx™. Only significant L-R pairs are shown, with SPP1 interacting exclusively with CD44. E. Heatmap illustrating the signal intensity of the sender and receiver in the SPP1 signaling pathway across all samples. F. Dot plots showing the outgoing communication patterns of secreting cells. Significant outgoing signals in the identified signaling pathways are represented by dots. G. Dot plots showing the incoming communication patterns of secreting cells. Significant incoming signals are represented by dots.

Article Snippet: Anti-SPP1 rabbit polyclonal antibody (HPA027541, Sigma-Aldrich, St. Louis, MO) and anti-CD44 mouse monoclonal antibody (UMAB134, OriGene Technologies, Rockville, MD) were used as primary antibodies.

Techniques: Protein-Protein interactions

Journal: bioRxiv

Article Title: Identification of SPP1-positive macrophages by single-cell spatial analysis in human lung tissues with mycobacterial infection

doi: 10.1101/2024.09.12.612778

Figure Lengend Snippet: SPP1 and CD44 expression in tissues from patients with granulomatous diseases. A. HE staining. B. Immunohistochemistry for SPP1. C. Immunofluorescence of SPP1 and CD44. Tissues from patients with sarcoidosis (left), granulomatosis with polyangiitis (GPA, middle), and rheumatoid nodule (RN, right) were investigated.

Article Snippet: Anti-SPP1 rabbit polyclonal antibody (HPA027541, Sigma-Aldrich, St. Louis, MO) and anti-CD44 mouse monoclonal antibody (UMAB134, OriGene Technologies, Rockville, MD) were used as primary antibodies.

Techniques: Expressing, Staining, Immunohistochemistry, Immunofluorescence