vegf  (Proteintech)

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

Journal: Bone & Joint Research

Article Title: OX40L in endothelial cells promotes temporomandibular joint subchondral bone angiogenesis and osteoclastogenesis in mice

doi: 10.1302/2046-3758.154.BJR-2025-0249.R1

Figure Lengend Snippet: Abnormal activation of OX40L in endothelial cells can promote angiogenesis and osteoclast induction. Human bone microvascular endothelial cells were pretreated with KY1005 for six hours, followed by treatment with sOX40 for 48 hours. a) Detection of vascular endothelial growth factor (VEGF) gene expression using quantitative polymerase chain reaction (qPCR). b) Detection of VEGF protein expression using western blotting. c) Representative images of endothelial cell scratch experiments. Scale bar: 100 µm. d) Representative images of endothelial cell tubule formation experiments. Scale bar: 100 µm. e) Representative image of tartrate-resistant acid phosphatase (TRAP) staining after endothelial cells were co-cultured with RAW264.7 cells. Scale bar: 50 µm. f) Bone resorption pits were shown by scanning electron microscopy. Scale bar: 500 nm.

Article Snippet: The membrane was blocked with 5% bovine serum albumin (Beyotime) and incubated overnight at 4°C with specific primary antibodies against vascular endothelial growth factor (VEGF) (1:1,000; Cat# 19003-1-AP, Proteintech, China) and glyceraldehyde 3-phosphate dehydrogenase (1:2,000; Cat# 60004-1-Ig, Clone 1E6D9, Proteintech).

Techniques: Activation Assay, Gene Expression, Real-time Polymerase Chain Reaction, Expressing, Western Blot, Staining, Cell Culture, Electron Microscopy

Journal: Bone & Joint Research

Article Title: OX40L in endothelial cells promotes temporomandibular joint subchondral bone angiogenesis and osteoclastogenesis in mice

doi: 10.1302/2046-3758.154.BJR-2025-0249.R1

Figure Lengend Snippet: a) Statistical analysis of VEGF protein expression levels. b) Statistical analysis of mobility in scratch experiments. c) and d) Statistical analysis of lumen number and main branch length in tubule formation experiments. e) to h) Detection of osteoprotegerin (OPG), receptor activator of nuclear factor κB ligand (RANKL), and macrophage colony-stimulating factor (M-CSF) secretion in endothelial cell supernatants by enzyme-linked immunosorbent assay (ELISA). i) Statistical analysis of resorption areas. Data are presented as the mean (SD) (n = 3). Statistical significance was determined by one-way analysis of variance followed by Tukey’s multiple comparison test. *p < 0.05, **p < 0.01, and ***p < 0.001.

Article Snippet: The membrane was blocked with 5% bovine serum albumin (Beyotime) and incubated overnight at 4°C with specific primary antibodies against vascular endothelial growth factor (VEGF) (1:1,000; Cat# 19003-1-AP, Proteintech, China) and glyceraldehyde 3-phosphate dehydrogenase (1:2,000; Cat# 60004-1-Ig, Clone 1E6D9, Proteintech).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Comparison