Journal: Advanced Science

Article Title: The Characteristics of Tumor Microenvironment by the Single Cell Profile in Endometrial Cancer of no Specific Molecular Profile (NSMP) Subtype

doi: 10.1002/advs.78332

Figure Lengend Snippet: Characterization of epithelial cells in NSMP EC. (A) Heatmap shows the CNV of epithelial cells in NSMP EC. (B) UMAP plot shows the distributions of different epithelial cell subtypes, color‐coded by subclusters. (C) Radar plot indicates the proportion of epithelial cell subtypes in NR and R tumors. (D) Box plot illustrates the stemness score of NR and R tumors at the cellular level. (E) UMAP plot shows the stemness score of different epithelial cell subtypes. (F) Heatmap shows 10 MPs of epithelial cells extracted by the NMF method. (G) Heatmap shows the expression of TFs in NR and R tumors. (H) Two‐sided bar graph displays the distribution of pathways in NR and R tumors. (I) Dot plot shows the expression of CD44 in epithelial cell subtypes. (J) Violin plot indicates the expression levels of CD44 from NR and R samples at the cellular level. (K) The spatial dot plots show the expression of CD44 in NR and R tumors. Dotted lines indicate the boundary of the tumor.

Article Snippet: The primary antibodies were: EPCAM (21050‐1‐AP, Proteintech), CD68 (25747‐1‐AP, Proteintech), SPP1 (22952‐1‐AP, Proteintech), CD4 (67786‐1‐lg, Proteintech) and CD44 (15675‐1‐AP, Proteintech).

Techniques: Expressing

Journal: Advanced Science

Article Title: The Characteristics of Tumor Microenvironment by the Single Cell Profile in Endometrial Cancer of no Specific Molecular Profile (NSMP) Subtype

doi: 10.1002/advs.78332

Figure Lengend Snippet: Macrophages with highly expressing SPP1 exhibit enhanced interactions with CD4+ T cells and epithelial cells expressing CD44 in R NSMP EC. (A) Chord diagram illustrates the SPP1 signaling in the R tumors. (B) Circle plots demonstrate the cell‐cell communications among Epi_C2, Epi_C3, macrophages, and CD4+ T cells. (C) The ligand‐receptor pair analysis among Epi_C2, Epi_C3, SPP1+ TAMs, and MKI67+ TAMs. SPP1‐CD44 axis emerged as the primary contributor to this interaction. (D) Representative images of mIHC staining for the SPP1+ CD68+ TAMs and CD44+ CD4+ T cells in NR and R NSMP EC. (E) The histogram illustrates the fraction of SPP1+ CD68+ TAMs in NR and R tumors. Data presented as mean ± SD. The p ‐value was calculated by Student's t‐test. (F) The spatial dot plots show the co‐localization of CD44 and SPP1 in NR and R tumors. Dotted lines indicate the boundary of the tumor. (G) The spatial position of SPP1+ CD68+ macrophages and CD44+ epithelial cells. White arrows indicate SPP1+ CD68+ macrophages; Orange arrows indicate SPP1‐ CD68+ macrophages. * p < 0.05.

Article Snippet: The primary antibodies were: EPCAM (21050‐1‐AP, Proteintech), CD68 (25747‐1‐AP, Proteintech), SPP1 (22952‐1‐AP, Proteintech), CD4 (67786‐1‐lg, Proteintech) and CD44 (15675‐1‐AP, Proteintech).

Techniques: Expressing, Analysis, Staining

Journal: Advanced Science

Article Title: The Characteristics of Tumor Microenvironment by the Single Cell Profile in Endometrial Cancer of no Specific Molecular Profile (NSMP) Subtype

doi: 10.1002/advs.78332

Figure Lengend Snippet: SPP1+ macrophages promote T cell stress and maintain epithelial cell stemness. (A, B) Western blot and RT‐qPCR results of SPP1 expression in macrophages with SPP1 knockdown. (C, D) Western blot and RT‐qPCR results of SPP1 expression in macrophages with overexpression of SPP1. (E) Western blot analysis of the protein expression of HSP70, LAG3, and PD‐1 in Jurkat cells after co‐culture with control and SPP1 knockdown macrophages. (F) qRT‐PCR analysis of the transcriptional level of HSPA1A and HSPA1B in Jurkat cells after co‐culture with control and SPP1 knockdown macrophages. (G) Western blot analysis of the protein expression of HSP70, LAG3, and PD‐1 in Jurkat cells after co‐culture with control, SPP1 overexpression, and SPP1 overexpression plus anti‐CD44 macrophages. (H) qRT‐PCR analysis of the transcriptional level of HSPA1A and HSPA1B in Jurkat cells after co‐culture with control, SPP1 overexpression, and SPP1 overexpression plus anti‐CD44 macrophages. (I) Western blot analysis of the protein expression of CD44 and CD133 in Ishikawa cells after co‐culture with control and SPP1 knockdown macrophages. (J) Western blot analysis of the protein expression of CD44 and CD133 in Ishikawa cells after co‐culture with control, SPP1 overexpression, and SPP1 overexpression plus anti‐CD44 macrophages. (K) Sphere formation assay was used to detect the stemness ability of Ishikawa cells after co‐culture with control and SPP1 knockdown macrophages. (L) A sphere formation assay was used to detect the stemness ability of Ishikawa cells after co‐culture with control, SPP1 overexpression, and SPP1 overexpression plus anti‐CD44 macrophages. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, ns: not significant.

Article Snippet: The primary antibodies were: EPCAM (21050‐1‐AP, Proteintech), CD68 (25747‐1‐AP, Proteintech), SPP1 (22952‐1‐AP, Proteintech), CD4 (67786‐1‐lg, Proteintech) and CD44 (15675‐1‐AP, Proteintech).

Techniques: Western Blot, Quantitative RT-PCR, Expressing, Knockdown, Over Expression, Analysis, Co-culture Assay, Control, Tube Formation Assay

Journal: Frontiers in Pharmacology

Article Title: Histopathology-guided spatial transcriptomic analysis identifies an ECM-associated stromal pattern in lacrimal gland adenoid cystic carcinoma with complementary proteomic and functional analyses

doi: 10.3389/fphar.2026.1949721

Figure Lengend Snippet: Bulk RNA-seq contextualizes spatially defined transcriptional programs in LGACC. (A) Bulk RNA-seq Transcriptomic Shifts. (B) Expression of representative receptor-related and ECM-associated genes in normal lacrimal gland and LGACC tissues. (C) Gene set enrichment analysis of spatially derived signatures in the bulk RNA-seq dataset. The Tumor Core-associated custom signature labeled “Spatial_Core_Survival” comprises MYB, MIF, CD44, CD74, and SDC4 and represents a spatially derived gene set. (D) ESTIMATE-derived ImmuneScore and StromalScore in normal lacrimal gland and LGACC samples.

Article Snippet: After blocking, the membranes were incubated overnight at 4 °C with primary antibodies against CD44 (1: 1000, Proteintech, 15675-1-AP) and β-actin (1: 2000, CST, 4970).

Techniques: RNA Sequencing, Expressing, Analysis, Derivative Assay, Labeling, Significance Assay

Journal: Frontiers in Pharmacology

Article Title: Histopathology-guided spatial transcriptomic analysis identifies an ECM-associated stromal pattern in lacrimal gland adenoid cystic carcinoma with complementary proteomic and functional analyses

doi: 10.3389/fphar.2026.1949721

Figure Lengend Snippet: Predicted region-to-region signaling interactions between histopathologically defined Tumor Core and Stroma Boundary compartments. (A) Global CellChat-inferred interaction network between the histopathologically defined Tumor Core and Stroma Boundary regions. (B) Bidirectional chord diagram showing candidate signaling pathways between the two annotated regions. (C) Predicted collagen-CD44-related interactions within the tumor-core region. (D) Predicted ECM-associated ligand-receptor interactions from the Stroma Boundary to the Tumor Core, including collagen- and fibronectin-related ligands with CD44 as a candidate receiving receptor.

Article Snippet: After blocking, the membranes were incubated overnight at 4 °C with primary antibodies against CD44 (1: 1000, Proteintech, 15675-1-AP) and β-actin (1: 2000, CST, 4970).

Techniques: Protein-Protein interactions, Significance Assay

Journal: Frontiers in Pharmacology

Article Title: Histopathology-guided spatial transcriptomic analysis identifies an ECM-associated stromal pattern in lacrimal gland adenoid cystic carcinoma with complementary proteomic and functional analyses

doi: 10.3389/fphar.2026.1949721

Figure Lengend Snippet: Spatial expression patterns of MIF, CD44, COL1A1, and COL1A2 in the analyzed LGACC section (A,B) Whole-tissue spatial expression maps of MIF and CD44, respectively. (C,D) COL1A1 and COL1A2 expression displayed within the manually annotated Tumor Core and Stroma Boundary regions.

Article Snippet: After blocking, the membranes were incubated overnight at 4 °C with primary antibodies against CD44 (1: 1000, Proteintech, 15675-1-AP) and β-actin (1: 2000, CST, 4970).

Techniques: Expressing, Tissue

Journal: Frontiers in Pharmacology

Article Title: Histopathology-guided spatial transcriptomic analysis identifies an ECM-associated stromal pattern in lacrimal gland adenoid cystic carcinoma with complementary proteomic and functional analyses

doi: 10.3389/fphar.2026.1949721

Figure Lengend Snippet: Type I collagen promotes SACC-LM cell spreading and migration. (A) Representative bright-field images showing the morphology of SACC-LM cells cultured on uncoated control plates (NC) or plates coated with type I collagen at 2 or 5 μg/cm 2 . (B) Quantification of the relative cell-free area from representative microscopic images. (C) CCK-8 assay showing cell viability at 0, 12, and 24 h. (D) Representative wound-healing images acquired at 0, 12, and 24 h after scratch generation. (E) Quantification of wound closure rates demonstrating enhanced migration in COL1-coated groups. (F) Representative images of migrated cells in the Transwell migration assay following crystal violet staining. (G) Quantification of migrated cell numbers showing significantly increased migration after COL1 coating. (H) Representative Western blot images showing CD44 protein expression in SACC-LM cells cultured on uncoated or COL1-coated plates. (I) Densitometric quantification of CD44 protein expression normalized to the loading control. * P < 0.05, ** P < 0.01.

Article Snippet: After blocking, the membranes were incubated overnight at 4 °C with primary antibodies against CD44 (1: 1000, Proteintech, 15675-1-AP) and β-actin (1: 2000, CST, 4970).

Techniques: Migration, Morphology, Cell Culture, Control, CCK-8 Assay, Transwell Migration Assay, Staining, Western Blot, Expressing

Journal: Frontiers in Pharmacology

Article Title: Histopathology-guided spatial transcriptomic analysis identifies an ECM-associated stromal pattern in lacrimal gland adenoid cystic carcinoma with complementary proteomic and functional analyses

doi: 10.3389/fphar.2026.1949721

Figure Lengend Snippet: CD44 knockdown attenuates COL1-induced wound closure in SACC-LM cells. (A) Representative Western blot showing efficient siRNA-mediated knockdown of CD44 in SACC-LM cells. β-actin was used as the loading control. (B) Representative wound-healing images of cells transfected with negative-control siRNA (si-NC), si-NC cultured on type I collagen-coated surfaces (si-NC + COL1), and CD44-targeting siRNA cultured on type I collagen-coated surfaces (si-CD44 + COL1) at 0, 12, and 24 h. Red lines indicate the wound margins. (C) Quantification of wound closure at 12 and 24 h. COL1 coating significantly increased wound closure in si-NC-transfected cells, whereas CD44 knockdown attenuated the COL1-induced increase in wound closure. * P < 0.05, ** P < 0.01.

Article Snippet: After blocking, the membranes were incubated overnight at 4 °C with primary antibodies against CD44 (1: 1000, Proteintech, 15675-1-AP) and β-actin (1: 2000, CST, 4970).

Techniques: Knockdown, Western Blot, Control, Transfection, Negative Control, Cell Culture

Journal: Journal of Dental Sciences

Article Title: Deer antler tip cell culture supernatant restores endothelial barrier function and angiotensin converting enzyme 2/endothelial nitric oxide synthase nitric oxide signaling impaired by Porphyromonas gingivalis lipopolysaccharide

doi: 10.1016/j.jds.2025.07.3521

Figure Lengend Snippet: Isolation and phenotypic characterization of primary cells from different regions of the velvet antler. (A) Representative photographs illustrate tissue sampling from Formosan sika deer during the velvet stage. Samples were collected from three anatomically defined regions tip, shaft, and burr as indicated in the schematic diagram. (B) Histological analyses using Masson’s trichrome and hematoxylin and eosin (H&E) staining reveal pronounced regional differences in tissue architecture. The tip region is characterized by loose connective tissue enriched with a dense microvascular network, accompanied by numerous hair follicles and sebaceous glands, consistent with an actively proliferative and angiogenic microenvironment. The shaft region exhibits increased tissue compactness with partially organized collagen fibers and reduced vascular density, indicating progressive differentiation and matrix remodeling. In contrast, the burr region displays tightly packed collagen bundles and early ossification fronts, reflecting a transition toward tissue maturation and mineralization. Scale bars: 200 μm. (C) Schematic workflow showing the process of primary cell isolation from freshly harvested antler tissues. After surface sterilization, tissues from the tip, shaft, and burr were minced and enzymatically digested (collagenase I + dispase II), followed by filtration, centrifugation, and primary culture. (D) Representative phase-contrast images of primary cells derived from each region. Tip-derived cells exhibit spindle-shaped, fibroblast-like morphology; shaft-derived cells show mixed elongated and polygonal forms; and burr-derived cells appear broad and flattened with partial mineralization features. (E) Immunofluorescence staining of cells for α-SMA, CD31, CD44, VEGFA, VEGFR, CD90, OCT4, and Slug, with DAPI nuclear counterstaining. Tip-derived cells express stemness-associated markers (CD44, CD90, OCT4, Slug); shaft-derived cells are enriched in endothelial and angiogenic markers (VEGFA, VEGFR, CD31); and burr-derived cells display fibroblastic and pre-osteogenic characteristics (α-SMA, VEGF/VEGFR). Scale bars = 100 μm. Abbreviations: DAPI, 4′,6-diamidino-2-phenylindole; α-SMA, alpha-smooth muscle actin; CD31, cluster of differentiation 31; CD44, cluster of differentiation 44; CD90, cluster of differentiation 90; OCT4, octamer-binding transcription factor 4; Slug, snail family transcriptional repressor 2 (SNAI2); VEGFA, vascular endothelial growth factor A; VEGFR, vascular endothelial growth factor receptor.

Article Snippet: The antibodies used included anti-CD44 antibody (15675-1-AP), anti-CD90 antibody (27178-1-AP), anti-OCT4/POU5F1 antibody (60242-1-Ig), anti-SNAI2/SLUG antibody (12129-1-AP), anti-VEGFA antibody (19003-1-AP), anti-VEGFR2/KDR antibody (26415-1-AP), anti-CD31 antibody (11265-1-AP), and anti-α-SMA antibody (67735-1-Ig), all purchased from Proteintech Group, Inc. (Rosemont, IL, USA).

Techniques: Isolation, Tissue, Sampling, Staining, Cell Isolation, Filtration, Centrifugation, Phase Contrast, Derivative Assay, Morphology, Immunofluorescence, Binding Assay, H&E Stain