vegfr2  (Cell Signaling Technology Inc)

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

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: Public GSE cohort analyses reveal conserved endothelial stress and remodeling programs in venous thrombosis. (A) Volcano plot and UMAP analysis of GSE103672 demonstrate clear transcriptomic separation between VTE and healthy controls. (B) Cross-cohort analysis of GSE19151 using volcano, radar, and bubble plots identifies recurrently amplified stress- and metabolism-related endothelial genes. (C) Heatmap of intersecting genes shared by both cohorts shows concordant expression patterns. (D) Pathway enrichment highlights endothelial metabolic regulation, redox responses, vascular remodeling, and VEGF/MAPK signaling associated with VTE. Abbreviations: ABCG2, ATP-binding cassette subfamily G member 2; ARL6IP5, ADP-ribosylation factor-like GTPase 6 interacting protein 5; CTL, control; CYP1B1, cytochrome P450 family 1 subfamily B member 1; ECM, extracellular matrix; GLO1, glyoxalase I; GNB1, G protein subunit beta 1; GO, Gene Ontology; GPX1, glutathione peroxidase 1; HINT1, histidine triad nucleotide-binding protein 1; HMGB1, high mobility group box 1; log2FC, log2 fold change; MAPK, mitogen-activated protein kinase; MMP10, matrix metalloproteinase 10; PGK1, phosphoglycerate kinase 1; PROCR, protein C receptor; PRDX4, peroxiredoxin 4; PRDX6, peroxiredoxin 6; R-HSA, Reactome pathway; RVTE, recurrent venous thromboembolism; S100A10, S100 calcium-binding protein A10; SOD1, superoxide dismutase 1; UMAP, uniform manifold approximation and projection; VEGFA, vascular endothelial growth factor A; VEGFR2, vascular endothelial growth factor receptor 2; VTE, venous thromboembolism; WP, WikiPathways.

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: Analysis, Amplification, Expressing, Binding Assay, Control

Journal: International Journal of Nanomedicine

Article Title: Levofloxacin-Catalase-PLGA Nanoparticles Potentiate Sonodynamic Therapy to Alleviate BCG-Induced Tuberculous Granulomas via PI3K/Akt/VEGF Pathway Suppression in Rats

doi: 10.2147/IJN.S628763

Figure Lengend Snippet: Microenvironmental features of 3D tuberculous spheroids. ( A ) Bright-field imaging of spheroid formation (scale bars: 360 μm (first three panels), 20 μm (right)); ( B ) Confocal immunofluorescence of HIF-1α and VEGF in control and tuberculous spheroids (scale bar: 50 μm); ( C ) Intracellular H 2 O 2 levels (n = 3); ( D and E ) qRT-PCR quantification of HIF-1α ( D ) and VEGF ( E ) mRNA expression in tuberculous spheroids and cell spheroids (n = 3); ( F and G ) Western blot ( F ) and grayscale analysis ( G ) of HIF-1α and VEGF protein levels by Image J (n = 3). *p < 0.05, **p < 0.01,* **p < 0.001.

Article Snippet: Membranes were incubated overnight at 4°C with primary antibodies against PI3K (CST, USA, 1:1000), p-PI3K (CST, USA, 1:1000), Akt (CST, USA, 1:1000), p-Akt (CST, USA, 1:1000), HIF-1α (Abcam, UK, 1:1000), VEGF (Proteintech, China, 1:1000), Bax (Proteintech, China, 1:1000), and Bcl-2 (Proteintech, China, 1:1500).

Techniques: Spheroids, Imaging, Confocal, Immunofluorescence, Control, Quantitative RT-PCR, Expressing, Western Blot, Analysis

Journal: International Journal of Nanomedicine

Article Title: Levofloxacin-Catalase-PLGA Nanoparticles Potentiate Sonodynamic Therapy to Alleviate BCG-Induced Tuberculous Granulomas via PI3K/Akt/VEGF Pathway Suppression in Rats

doi: 10.2147/IJN.S628763

Figure Lengend Snippet: US + LEV@CAT-NPs treatment downregulates HIF-1α/VEGF expression in BCG-induced tuberculous spheroids. ( A and B ) Immunofluorescence images of HIF-1α ( A ) and VEGF ( B ) in tuberculous spheroids following various treatments; scale bar: 100 μm. ( C and D ) Relative mRNA levels of HIF-1α and VEGF in tuberculous spheroids across different groups (n = 3). ( E ) Secreted VEGF concentration in the culture supernatant of tuberculous spheroids (n = 3).* **P < 0.001.

Article Snippet: Membranes were incubated overnight at 4°C with primary antibodies against PI3K (CST, USA, 1:1000), p-PI3K (CST, USA, 1:1000), Akt (CST, USA, 1:1000), p-Akt (CST, USA, 1:1000), HIF-1α (Abcam, UK, 1:1000), VEGF (Proteintech, China, 1:1000), Bax (Proteintech, China, 1:1000), and Bcl-2 (Proteintech, China, 1:1500).

Techniques: Expressing, Spheroids, Immunofluorescence, Concentration Assay

Journal: International Journal of Nanomedicine

Article Title: Levofloxacin-Catalase-PLGA Nanoparticles Potentiate Sonodynamic Therapy to Alleviate BCG-Induced Tuberculous Granulomas via PI3K/Akt/VEGF Pathway Suppression in Rats

doi: 10.2147/IJN.S628763

Figure Lengend Snippet: Transcriptomic and functional validation that US + LEV@CAT-NPs suppress VEGF expression via PI3K/Akt axis inhibition. ( A ) KEGG enrichment plot from RNA-seq analysis identifying the PI3K/Akt pathway as one of the top enriched pathways; ( B and C ) Western blot analysis ( B ) and its quantitative results ( C ) confirming the downregulation of HIF-1α, VEGF, and reduced phosphorylation levels of PI3K and Akt; ( D and E ) Pharmacological inhibition with LY294002 demonstrating that VEGF suppression is dependent on PI3K/Akt signaling, as shown by representative blots ( D ) and quantitative analysis of p-PI3K/PI3K, p-Akt/Akt ratio and secreted VEGF levels ( E ). * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: Membranes were incubated overnight at 4°C with primary antibodies against PI3K (CST, USA, 1:1000), p-PI3K (CST, USA, 1:1000), Akt (CST, USA, 1:1000), p-Akt (CST, USA, 1:1000), HIF-1α (Abcam, UK, 1:1000), VEGF (Proteintech, China, 1:1000), Bax (Proteintech, China, 1:1000), and Bcl-2 (Proteintech, China, 1:1500).

Techniques: Functional Assay, Biomarker Discovery, Expressing, Inhibition, RNA Sequencing, Analysis, Western Blot, Phospho-proteomics

Journal: International Journal of Nanomedicine

Article Title: Levofloxacin-Catalase-PLGA Nanoparticles Potentiate Sonodynamic Therapy to Alleviate BCG-Induced Tuberculous Granulomas via PI3K/Akt/VEGF Pathway Suppression in Rats

doi: 10.2147/IJN.S628763

Figure Lengend Snippet: Conditioned medium from US + LEV@CAT-NPs-treated tuberculous spheroids inhibits angiogenesis in HUVECs.( A ) Schematic diagram of the experimental workflow for evaluating the effects of conditioned media (CM) from US + LEV@CAT-NPs-treated tuberculous spheroids on HUVEC angiogenesis; ( B ) Representative images of tube formation by HUVECs cultured with different CM; scale bar: 50 μm; ( C ) Immunofluorescence staining of VEGF in HUVECs; scale bar: 20 μm; ( D and E ) Quantitative analysis of total tube length ( D ) and coverage area ( E ) for HUVEC tube‑formation assays (n = 3). Both tube length and coverage area were normalized to the CM‑Control group (set as 100%). Data are shown as mean ± SD. ( F ) qRT-PCR analysis of VEGF mRNA levels in HUVECs (n = 3). * P < 0.05, *** P < 0.001.

Article Snippet: Membranes were incubated overnight at 4°C with primary antibodies against PI3K (CST, USA, 1:1000), p-PI3K (CST, USA, 1:1000), Akt (CST, USA, 1:1000), p-Akt (CST, USA, 1:1000), HIF-1α (Abcam, UK, 1:1000), VEGF (Proteintech, China, 1:1000), Bax (Proteintech, China, 1:1000), and Bcl-2 (Proteintech, China, 1:1500).

Techniques: Spheroids, Cell Culture, Immunofluorescence, Staining, Analysis, Quantitative RT-PCR

Journal: International Journal of Nanomedicine

Article Title: Levofloxacin-Catalase-PLGA Nanoparticles Potentiate Sonodynamic Therapy to Alleviate BCG-Induced Tuberculous Granulomas via PI3K/Akt/VEGF Pathway Suppression in Rats

doi: 10.2147/IJN.S628763

Figure Lengend Snippet: Effects of different treatments on vascular structure and function, hypoxic microenvironment, and related signaling molecules in tuberculous granulomas in vivo. ( A ) Vascular density and maturity (pericyte coverage) assessed by immunofluorescence co-localization of α-SMA (green) and CD31 (red); scale bar: 50 μm; ( B ) Vascular leakage assessed by co-staining of 70 kDa FITC-dextran (green) and CD31 (red); scale bar: 100 μm; ( C ) Vascular perfusion assessed by co-localization of FITC-lectin (green) and CD31 (red); scale bar: 25 μm; ( D ) Immunohistochemical staining of HIF-1α; scale bar: 40 μm; ( E ) Immunofluorescence detection of hypoxic regions using pimonidazole (PIMO);scale bar: 100 μm; ( F ) Analysis of oxygen saturation (sO 2 ) in granulomas by photoacoustic imaging. Left panel: Representative photoacoustic imaging of granulomas; Right panel: Quantitative comparison of sO 2 levels in granulomas between the Control and US + LEV@CAT-NPs groups. ( G ) Semi-quantitative analysis of PIMO + reas in ( E ). ( H ) Immunohistochemical staining images of VEGF and VEGFR2; scale bar: 40 μm. ***P<0.001 .

Article Snippet: Membranes were incubated overnight at 4°C with primary antibodies against PI3K (CST, USA, 1:1000), p-PI3K (CST, USA, 1:1000), Akt (CST, USA, 1:1000), p-Akt (CST, USA, 1:1000), HIF-1α (Abcam, UK, 1:1000), VEGF (Proteintech, China, 1:1000), Bax (Proteintech, China, 1:1000), and Bcl-2 (Proteintech, China, 1:1500).

Techniques: In Vivo, Immunofluorescence, Staining, Contrast Agent Perfusion, Immunohistochemical staining, Analysis, Imaging, Comparison, Control