recombinant human vegf rhvegf  (R&D Systems)

Journal: Annals of Hematology

Article Title: Reduced number of endothelial progenitor cells in adult patients with beta thalassemia major

doi: 10.1007/s00277-026-06979-1

Figure Lengend Snippet: Violin plot representations of the proportion of ( a ) VEGFR-2 + CD34 + cells and ( b ) VEGFR-2 + CD133 + cells in TM patients (“TM”) ( n = 25) vs. healthy controls (“Controls”) ( n = 11)

Article Snippet: Briefly: isolated cells were stained with FITC-labeled VEGFR-2 (vascular endothelial growth factor receptor-2), a key endothelial lineage marker (R&D, Minneapolis, USA), CY5.5 labeled CD45 (Dako, Denmark), and either PE labeled CD133 or CD34 (Miltenyi Biotech, Auburn, CA, USA).

Techniques:

Journal: Annals of Hematology

Article Title: Reduced number of endothelial progenitor cells in adult patients with beta thalassemia major

doi: 10.1007/s00277-026-06979-1

Figure Lengend Snippet: Violin plot showing the proportion of VEGFR-2 + CD34 + cells in TM patients with a history of thromboembolic events ( n = 6) vs. TM patients with no history of thromboembolic events ( n = 19)

Article Snippet: Briefly: isolated cells were stained with FITC-labeled VEGFR-2 (vascular endothelial growth factor receptor-2), a key endothelial lineage marker (R&D, Minneapolis, USA), CY5.5 labeled CD45 (Dako, Denmark), and either PE labeled CD133 or CD34 (Miltenyi Biotech, Auburn, CA, USA).

Techniques:

Journal: Frontiers in Immunology

Article Title: Dysregulated hyaluronan metabolism drives inflammation and angiogenesis in proliferative diabetic retinopathy

doi: 10.3389/fimmu.2026.1724199

Figure Lengend Snippet: Expression levels of hyaluronidase (Hyal)-1 (A) , CD44 (B) and receptor for hyaluronan-mediated motility (RHAMM) (C) in the retinal lysates of non-diabetic control rats (C) (n=12) and diabetic rats (D) (n=12) were determined by Western blot analysis. After determination of the intensity of the protein bands, intensities were adjusted to those of β-actin in the samples. Oxidative stress was monitored with the use of 2’,7’-Dichlorofluorescein (DCF) fluorescence intensity analysis (D) . Results are expressed as mean ± standard deviation. Ultra-Low molecular weight hyaluronan (ULMW-HA) induces breakdown of blood-retinal barrier (E) . ULMW-HA was injected intravitreally at the dose of 50 ng in 5 µL in one eye and the same volume of phosphate-buffered saline (PBS) was injected in the contralateral eye of normal rats. The BRB was quantified with the fluorescein isothiocyanate-conjugated dextran technique. Results are expressed as mean ± standard deviation of 12 rats. *p < 0.05 compared to the values obtained from PBS-injected eyes. (independent t-test). Western blot analysis of retinas demonstrated that intravitreal injection of ULMW-HA induced significant upregulation of the expression of phospho-NF-κB (F) , phospho-ERK1/2 (G) , vascular endothelial growth factor (VEGF) (H) , intercellular adhesion molecule-1 (ICAM-1) (I) , vascular cell adhesion molecule-1 (VCAM-1) (J) and high-mobility group box-1 (HMGB1) (K) . Results are expressed as mean ± standard deviation or standard error of mean of 8–10 rats in each group (*p < 0.05; independent t-test).

Article Snippet: To determine the presence of Hyal-1, Hyal-2, HAS2, CD44, syndecan-1, heparan sulphate and RHAMM in the vitreous samples, equal volumes (10 μL) of vitreous samples were boiled in Laemmli’s sample buffer (1:1, v/v) under reducing condition for 10 min. Immunodetection was performed with the use of rabbit polyclonal anti-Hyal-1 antibody (1:1000, NBP2-16906, Novus Biologicals), mouse polyclonal anti-Hyal-2 antibody (1:1000, H00008692-B02P, Novus Biologicals), mouse monoclonal anti-HAS2 antibody (1:1000, ab140671, Abcam), rabbit monoclonal anti-CD44 antibody (1:1000, ab189524, Abcam), rabbit monoclonal anti-RHAMM antibody (1:1000, ab124729, Abcam), rabbit monoclonal anti-phospho-extracellular signal-regulated kinase (ERK)1/2 antibody (1:1000, MAB1018, R&D Systems), rabbit polyclonal anti-p65 subunit of nuclear factor-kappa B (phospho-NF-κB) (1:1000, NB100-82086, Novus Biologicals), rabbit polyclonal anti-high-mobility group box1 (HMGB1) (1:1000, Cat. no. ab18256, Abcam), mouse monoclonal anti-VEGF antibody (1:750, MAB293, R&D Systems), mouse monoclonal anti-intercellular adhesion molecule-1 (ICAM-1) antibody (1:100, sc-8439, Santa Cruz Biotechnology Inc.), and mouse monoclonal anti-vascular cell adhesion molecule-1 (VCAM-1) antibody (1:100, sc-13160, Santa Cruz Biotechnology Inc.).

Techniques: Expressing, Control, Western Blot, Fluorescence, Standard Deviation, Molecular Weight, Injection, Saline

Journal: Frontiers in Immunology

Article Title: Dysregulated hyaluronan metabolism drives inflammation and angiogenesis in proliferative diabetic retinopathy

doi: 10.3389/fimmu.2026.1724199

Figure Lengend Snippet: Human retinal Müller glial cells were left untreated or treated with ultra-low molecular weight hyaluronan (ULMW-HA) (50 µg/mL) for 24 (h) (A) Protein expression of phospho-ERK1/2 and phospho-NFκB in cell lysates was determined by Western blot analysis. Levels of high mobility group box-1 (HMGB1) were quantified in the culture media by ELISA. Results are expressed as mean ± standard deviation from three different experiments each performed in triplicate (*p < 0.05; independent t-test). (B) Human retinal Müller glial cells were left untreated or treated with ULMW-HA, ULMW-HA plus BAY11-7085 (5 µM) or (C) ULMW-HA plus U-0126 (5 µM). Levels of vascular endothelial growth factor (VEGF), angiopoietin and monocyte chemotactic protein-1 (MCP-1/CCL2) were quantified in the culture media by ELISA. Results are expressed as mean ± standard deviation or standard error of mean from three different experiments each performed in triplicate. One-way ANOVA and independent t-test were used for comparisons between three groups and two groups, respectively. *p < 0.05 compared with values obtained from untreated cells; #p < 0.05 compared with ULMW-HA plus BAY11–7085 or U-0126 treated cells. (D, E) Human retinal Müller glial cells were left untreated or treated with high glucose (HG) (25 mM), cobalt chloride (CoCl 2 ) (300 µM) or tumor necrosis factor-α (TNF-α) (5 ng/mL) with or without apigenin (10 µg/mL) for 24 (h) For HG treatment, cultures containing 25 mM mannitol were used as a control. Levels of monocyte chemotactic protein-1 (MCP-1/CCL2) (D) and vascular endothelial growth factor (VEGF) (E) were quantified in the culture media by ELISA. The results are expressed as mean ± standard deviation from three different experiments each performed in triplicate. One-way ANOVA and independent t-test were used for comparisons between three and two groups, respectively. *p < 0.05 compared with values obtained from control cells. #p < 0.05 compared with values obtained from stimulated cells.

Article Snippet: To determine the presence of Hyal-1, Hyal-2, HAS2, CD44, syndecan-1, heparan sulphate and RHAMM in the vitreous samples, equal volumes (10 μL) of vitreous samples were boiled in Laemmli’s sample buffer (1:1, v/v) under reducing condition for 10 min. Immunodetection was performed with the use of rabbit polyclonal anti-Hyal-1 antibody (1:1000, NBP2-16906, Novus Biologicals), mouse polyclonal anti-Hyal-2 antibody (1:1000, H00008692-B02P, Novus Biologicals), mouse monoclonal anti-HAS2 antibody (1:1000, ab140671, Abcam), rabbit monoclonal anti-CD44 antibody (1:1000, ab189524, Abcam), rabbit monoclonal anti-RHAMM antibody (1:1000, ab124729, Abcam), rabbit monoclonal anti-phospho-extracellular signal-regulated kinase (ERK)1/2 antibody (1:1000, MAB1018, R&D Systems), rabbit polyclonal anti-p65 subunit of nuclear factor-kappa B (phospho-NF-κB) (1:1000, NB100-82086, Novus Biologicals), rabbit polyclonal anti-high-mobility group box1 (HMGB1) (1:1000, Cat. no. ab18256, Abcam), mouse monoclonal anti-VEGF antibody (1:750, MAB293, R&D Systems), mouse monoclonal anti-intercellular adhesion molecule-1 (ICAM-1) antibody (1:100, sc-8439, Santa Cruz Biotechnology Inc.), and mouse monoclonal anti-vascular cell adhesion molecule-1 (VCAM-1) antibody (1:100, sc-13160, Santa Cruz Biotechnology Inc.).

Techniques: Molecular Weight, Expressing, Western Blot, Enzyme-linked Immunosorbent Assay, Standard Deviation, Control

Journal: Frontiers in Immunology

Article Title: Dysregulated hyaluronan metabolism drives inflammation and angiogenesis in proliferative diabetic retinopathy

doi: 10.3389/fimmu.2026.1724199

Figure Lengend Snippet: Human retinal microvascular endothelial cells (HRMECs) were left untreated or treated with high glucose (HG) (25 mM) (A) , cobalt chloride (CoCl 2 ) (300 µM) (B) or tumor necrosis factor-α (TNF-α) (5 ng/mL) (C) with or without apigenin (10 µg/mL). For HG treatment, cultures treated with mannitol (25 mM) were used as a control. Levels of soluble syndecan-1 were quantified in the culture media by ELISA. Results are expressed as mean ± standard deviation from three different experiments each performed in triplicate. One-way ANOVA and independent t-test were used for comparisons between three and two groups, respectively. *p < 0.05 compared with values obtained from control cells. #p < 0.05 compared with values obtained from cells treated with HG, CoCl 2 or TNF-α. HRMECs were left untreated or were stimulated with ultra-low molecular weight – hyaluronan (ULMW-HA) (50 µg/mL) for 24 (h). Protein expression of phospho-ERK1/2 in the cell lysates was determined by Western blot analysis (D) . Results are expressed as mean ± standard deviation from three different experiments each performed in triplicate (*p < 0.05; independent t-test). A scratch was performed in confluent monolayers of overnight starved HRMECs with a micropipette tip subsequently, the cultures were left untreated or treated either with VEGF (10 ng/mL) or with ULMW-HA (100 µg/mL) for 16 (h) Cells were visualized using an inverted microscope. Two independent experiments were performed. Each experiment was done in duplicate, and 2–3 independent field images were taken for the migration analysis which was done by using Image J software. In the Figure, one representative image is illustrated, and the bar graphs show the analysis of all the images from each group represented as fold-change in migration versus control (E) . Results are expressed as mean ± standard deviation. One-way ANOVA and independent t-test were used for comparisons between three and two groups, respectively. *p < 0.05 compared with values obtained from control cells.

Article Snippet: To determine the presence of Hyal-1, Hyal-2, HAS2, CD44, syndecan-1, heparan sulphate and RHAMM in the vitreous samples, equal volumes (10 μL) of vitreous samples were boiled in Laemmli’s sample buffer (1:1, v/v) under reducing condition for 10 min. Immunodetection was performed with the use of rabbit polyclonal anti-Hyal-1 antibody (1:1000, NBP2-16906, Novus Biologicals), mouse polyclonal anti-Hyal-2 antibody (1:1000, H00008692-B02P, Novus Biologicals), mouse monoclonal anti-HAS2 antibody (1:1000, ab140671, Abcam), rabbit monoclonal anti-CD44 antibody (1:1000, ab189524, Abcam), rabbit monoclonal anti-RHAMM antibody (1:1000, ab124729, Abcam), rabbit monoclonal anti-phospho-extracellular signal-regulated kinase (ERK)1/2 antibody (1:1000, MAB1018, R&D Systems), rabbit polyclonal anti-p65 subunit of nuclear factor-kappa B (phospho-NF-κB) (1:1000, NB100-82086, Novus Biologicals), rabbit polyclonal anti-high-mobility group box1 (HMGB1) (1:1000, Cat. no. ab18256, Abcam), mouse monoclonal anti-VEGF antibody (1:750, MAB293, R&D Systems), mouse monoclonal anti-intercellular adhesion molecule-1 (ICAM-1) antibody (1:100, sc-8439, Santa Cruz Biotechnology Inc.), and mouse monoclonal anti-vascular cell adhesion molecule-1 (VCAM-1) antibody (1:100, sc-13160, Santa Cruz Biotechnology Inc.).

Techniques: Control, Enzyme-linked Immunosorbent Assay, Standard Deviation, Molecular Weight, Expressing, Western Blot, Inverted Microscopy, Migration, Software

Journal: Lab on a Chip

Article Title: Label-free assessment of a microfluidic vessel-on-chip model with visible-light optical tomography reveals structural changes in vascular networks

doi: 10.1039/d5lc00927h

Figure Lengend Snippet: Time-resolved imaging of vascular network development in vessel-on-chip by optical coherence tomography. A) Timeline of disease modeling. Vessel-on-chips were loaded and vessels were allowed to grow for 2 days. Thereafter, the vascular network was subjected to control medium, medium with high glucose and added TNF-α and IL-6, and VEGF medium for 3 more days. Vessel-on-chips were measured every day after day 2. B) Minimum intensity projections, showing the change in the vascular network in the control condition over the course of 5 days. C) Minimum intensity projections, displaying changes in vascular network for the high glucose condition on day 4 and 5. D) Minimum intensity projections, exhibiting changes in the vascular network for the VEGF condition on day 4 and 5. For a full overview of the process, see Fig. S2 in SI. Representative images shown, scale bar = 500 μm.

Article Snippet: On day 3, vascular specification was induced by adding 50 ng ml −1 vascular endothelial growth factor (VEGF) (Miltenyi Biotec, Germany) and 10 μM SB431542 (Tocris Bioscience, UK) in BPEL medium to the cells.

Techniques: Imaging, Tomography, Control

Journal: Lab on a Chip

Article Title: Label-free assessment of a microfluidic vessel-on-chip model with visible-light optical tomography reveals structural changes in vascular networks

doi: 10.1039/d5lc00927h

Figure Lengend Snippet: Change in vessel thickness in the vessel-on-chip over the treatment period, for the different treatments. A) Control condition on day 2 to 5, B) high glucose with added TNF-α and IL-6 condition on day 4 to 5, and C) VEGF treatment on day 4 to 5. For a full overview of the process, see Fig. S3 in SI. Representative images shown, scale bar = 500 μm.

Article Snippet: On day 3, vascular specification was induced by adding 50 ng ml −1 vascular endothelial growth factor (VEGF) (Miltenyi Biotec, Germany) and 10 μM SB431542 (Tocris Bioscience, UK) in BPEL medium to the cells.

Techniques: Control

Journal: Lab on a Chip

Article Title: Label-free assessment of a microfluidic vessel-on-chip model with visible-light optical tomography reveals structural changes in vascular networks

doi: 10.1039/d5lc00927h

Figure Lengend Snippet: Overlay of the variation in the number of branches and vessel length under the different conditions during treatment of the vessel-on-chip for A) the control condition on day 2 to 5, B) the high glucose with added TNF-α and IL-6 condition on day 4 and 5, and C) the VEGF condition on day 4 to 5. For a full overview of the process, see Fig. S4 in SI. Representative images shown, scale bar = 500 μm.

Article Snippet: On day 3, vascular specification was induced by adding 50 ng ml −1 vascular endothelial growth factor (VEGF) (Miltenyi Biotec, Germany) and 10 μM SB431542 (Tocris Bioscience, UK) in BPEL medium to the cells.

Techniques: Control

Journal: Lab on a Chip

Article Title: Label-free assessment of a microfluidic vessel-on-chip model with visible-light optical tomography reveals structural changes in vascular networks

doi: 10.1039/d5lc00927h

Figure Lengend Snippet: Quantitative properties of the vascular network during treatment for all conditions. A) Vascularity index (VI), B) mean thickness, C) total vessel length, and D) number of branching points. Data are presented in boxplots from four individual microfluidic chips ( n = 4). Statistical analyses were performed using one-way ANOVA followed by a Student's t -test. * indicates p < 0.05. E) Minimum intensity projections from Fig. S2 showing the change in the vascular network in the control, high glucose with added TNF-α and IL-6, and VEGF condition over the course of 5 days. Representative images shown, scale bar = 500 μm.

Article Snippet: On day 3, vascular specification was induced by adding 50 ng ml −1 vascular endothelial growth factor (VEGF) (Miltenyi Biotec, Germany) and 10 μM SB431542 (Tocris Bioscience, UK) in BPEL medium to the cells.

Techniques: Control

Journal: Nature

Article Title: GlycoRNA complexed with heparan sulfate regulates VEGF-A signalling

doi: 10.1038/s41586-025-10052-8

Figure Lengend Snippet: a , Quantification of 10E4, Siglec-11, 9D5, cs-DDX21 and cs-hnRNP-U intensity per cell from three independent staining experiments on HUVECs. Data are mean ± s.e.m. a.u., arbitrary units. b , Western blot analysis of whole-cell lysates isolated from HUVECs after starvation and treatment with an RNase pool followed by 3 ng ml −1 VEGF-A 165 , VEGF-A 121 or EGF stimulation (top). Quantification of the ratio of pERK to total ERK was also calculated across the biological triplicates (bottom). Statistical assessment was performed with a two-sided Student’s t -test and P values are shown. Data are mean ± s.e.m. c , Representative images of starved HUVECs treated with an RNase pool, and subsequently with VEGF-A 165 or VEGF-A 121 , finally stained with anti-VEGF-A 165 (red) or anti-VEGF-A (red). Statistical assessment was performed with a Student’s t -test and P values are shown. Data are mean ± s.e.m. NS, not significant. d , Representative images of HUVECs treated with an RNase pool and stained with anti-VEGFR2 (red). Statistical assessment was performed with a Student’s t -test and P values are shown. Data are mean ± s.e.m. e , Representative images of starved HUVECs treated with VEGF-A 165 and then co-stained with anti-VEGF-A 165 (purple) and Siglec-11 (yellow). Three independent experiments were performed. f , Schematic of the microfluidic chip (top left) used to grow HUVECs without (top middle) and with (top right) RNase A for 6 days. Representative images of BFP expressed in the HUVECs. Statistical assessment of the total migration area was performed using an unpaired two-sided Student’s t -test (bottom). Data are mean ± s.e.m. Four independent experiments were performed. g , Representative image (maximum z -projection view (left) and z -projection slice view (right)) of sprouts from a +RNase A device for the cells (blue), F-actin (red) and PECAM1 (green). Four independent experiments were performed.

Article Snippet: Anti-VEGF-A 165 (AF293-NA, R&D Systems; immunoblot 1:1,000) and donkey anti-goat IgG secondary antibody (92632214, LI-COR Biosciences; immunoblot 1:1,000) were used as primary and secondary antibodies, separately.

Techniques: Staining, Western Blot, Isolation, Migration

Journal: Nature

Article Title: GlycoRNA complexed with heparan sulfate regulates VEGF-A signalling

doi: 10.1038/s41586-025-10052-8

Figure Lengend Snippet: a . Western blot analysis of whole cell lysate isolated from HUVECs after starvation and treatment with RNase pool followed by 25 ng/mL VEGF-A 165 stimulation. Quantification of the ratio of phosphorylated ERK (pERK) to total ERK is calculated across the biological triplicates. Statistical assessment was performed with a two-sided Student's t -test and P values are shown. Data are mean ± s.e.m. b . Western blot analysis as in (A) with HUVECs stimulated with 25 ng/mL of VEGF-A 121 . Statistical assessment and data are as in ( a ). c . Western blot analysis as in (A) with HUVECs stimulated with 25 ng/mL of EGF. Statistical assessment and data are as in ( a ). d . Western blot analysis as in (A) with HUVECs stimulated with 3 ng/mL of VEGF-A 165 . Quantification of the ratio of phosphorylated VEGFR2 (pVEGFR2) to total VEGFR2 is calculated across the biological triplicates. Statistical assessment and data are as in ( a ). e . MST assay of VEGF-A and small RNA binding. n = 3 biologically independent repeats. Data are mean ± s.e.m. f . Representative images of HUVECs after serum starvation and treated with or without VEGF-A 165 and then co-stained with anti-VEGF-A 165 (purple) and Siglec-11 (yellow). Zoomed region shown as an inset. Scale bar, 10 µm. 3 independent experiments were performed. g . Nearest neighbor distance analysis of the VEGF-A 165 and Siglec-11 in Figure S5E. For each pair, the nm distance from VEGF-A 165 to Siglec-11 was calculated across. These values were plotted in a density histogram. h . The four biological replicate microfluidic chip without (left) and with (right) 10 µM RNase A for 6 days. Scale bar, 200 µm. i . Immunofluorescence confocal image (maximum z-projection view) of HUVEC structures after 6 days of +RNase treatment on microfluidic chip. Cells (blue), F-actin (red), and PECAM1 (green). Scale bar, 100 µm. 4 independent experiments were performed.

Article Snippet: Anti-VEGF-A 165 (AF293-NA, R&D Systems; immunoblot 1:1,000) and donkey anti-goat IgG secondary antibody (92632214, LI-COR Biosciences; immunoblot 1:1,000) were used as primary and secondary antibodies, separately.

Techniques: Western Blot, Isolation, RNA Binding Assay, Staining, Immunofluorescence

Journal: Nature

Article Title: GlycoRNA complexed with heparan sulfate regulates VEGF-A signalling

doi: 10.1038/s41586-025-10052-8

Figure Lengend Snippet: a . Western blot analysis of the indicated amount of VEGF-A 165 . 2 independent experiments were performed. b . Lysates from HUVECs after serum starvation, treatment with or without 25 ng/mL VEGF-A 165 , and UV-crosslinking, were treated with or without RNase pool and immunoprecipitated (IP) with an anti-VEGF-A antibody (Proteintech). Immunoprecipitated samples were analyzed by Western blot using an anti-VEGF-A 165 antibody (R&D system). 3 independent experiments were performed. c . Principal component analysis (PCA) of VEGF-A 165 RIP-seq results. d . Enriched transcripts in VEGF-A 165 RIP-seq. Each dot represents a unique small non-coding RNA (ncRNA) transcript, colored by biotype as indicated in the legend. The red dotted line marks the filtering citeria: log 2 FoldChange (IP/Input) > 0.5 and adjusted p-value < 0.05. Statistical assessment was performed with a two-sided Wald test and P values are adjusted for multiple comparisons using the Benjamini-Hochbery procedure. Solid circles indicate true positive hits that were also significantly enriched relative to the IgG control (see for details). e . Abundance of enriched transcripts grouped by biotype. Color scheme matches ( d ). f . Overlap between enriched transcripts and glycoRNA defined by ManNAz-seq, grouped by ncRNA family. The p-value was calculated using a two-sided hypergeometric test with all human small ncRNAs as the background. g . Western blot analysis of the beads pre-conjugated with 5 µg of VEGF-A 165 . h . Representative images of the indicated HUVECs stained with anti-VEGF-A 165 (red). Scale bar, 10 µm. Quantification of the images with number of cells noted per biological triplicate. Statistical assessment was performed with a two-sided Student's t -test and P values are shown. Data are mean ± s.e.m. i . Western blot analysis of whole cell lysate isolated from the indicated HUVECs. Quantification of the ratio of phosphorylated VEGFR2 (pVEGFR2) to total VEGFR2 is calculated across the biological triplicates. Statistical assessment was performed with a two-sided Student's t -test and P values are shown. Data are mean ± s.e.m. j . In vitro IP of VEGF-A 165 HS WT or HS(R/K) with small RNA and rPAL. 3 independent experiments were performed. k . MST assay of VEGF-A and small RNA treated with sialidase binding. n = 3 biologically independent repeats. Data are mean ± s.e.m. l . RNA-seq analysis of the sulfotransferases expressed from HUVECs and their FPKM values from ref. . m . EMSA analysis of the indicated VEGF-A proteins with or without the addition of rHS29.

Article Snippet: Anti-VEGF-A 165 (AF293-NA, R&D Systems; immunoblot 1:1,000) and donkey anti-goat IgG secondary antibody (92632214, LI-COR Biosciences; immunoblot 1:1,000) were used as primary and secondary antibodies, separately.

Techniques: Western Blot, Immunoprecipitation, Control, Staining, Isolation, In Vitro, Binding Assay, RNA Sequencing