human vascular endothelial growth factor 165  (R&D Systems)

Journal: bioRxiv

Article Title: The LIF-LIFR Axis Promotes Liver Regeneration via Modulation of Angiogenesis and HGF Release from LSECs

doi: 10.64898/2026.02.24.707802

Figure Lengend Snippet: (A-C) Proliferation assays of primary LSECs treated with increasing doses of hLIF (A), hOSM (B) or hCT-1 (C). 100 ng/mL hLIF was used as positive control. (D) Proliferation assays of LSECs isolated from lifrΔEC mice treated with increasing doses of hLIF. (E&F) Proliferation assays of LSECs isolated from lifr f/f (E) or lifrΔEC (F) mice treated with increasing doses of hVEGF. (G&H) Western blot analysis of primary LSECs treated with hVEGF (G) or hLIF (H) (I) Western blot of primary hepatocytes treated with hLIF. (J) Proliferation assays of primary hepatocytes treated with increasing doses of hLIF. (K) Proliferation assays of AML12 cells treated with increasing doses of hLIF. (L) Immunofluorescence staining of AML12 cells treated with 80 ng/mL hHGF or hLIF. DAPI (blue), Ki67 (green). Scale bar: 50 μm.

Article Snippet: The proteins which included hLIF (Abcam, #ab287941), hVEGF (R&D system, #293-VE), hOSM (R&D system, #8475-OM), hCT1 (R&D system, 612-CD-010), IL-6 (PeproTech, 200-06), hHGF(R&D system, 294-HG), mHGF (R&D system, 2207-HG-025), EGF (R&D system, 236-EG), aLIF (UCSD), MAb mouse IgG2a isotype control (BioXcell,C1.18.4, #BE0085), anti-VEGF antibody B20-4.1.1 (Genentech) were added to cells at various concentrations, as indicated in the figures.

Techniques: Positive Control, Isolation, Western Blot, Immunofluorescence, Staining

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: Enzyme-linked immunosorbent assay (ELISA) kits for human Hyal-1 (Cat No DY7358), human Hyaluronan (Cat No DY3614-05), and human syndecan-1 (Cat No DY2780), human vascular endothelial growth factor (VEGF) (Cat No DY293B), human monocyte chemoattractant protein-1 (MCP-1/CCL2) (Cat No DY279), human matrix metalloproteinase-9 (MMP-9) (Cat No DY911) and human angiopoietin-2 (Cat No DY623) were purchased from R&D Systems.

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: Enzyme-linked immunosorbent assay (ELISA) kits for human Hyal-1 (Cat No DY7358), human Hyaluronan (Cat No DY3614-05), and human syndecan-1 (Cat No DY2780), human vascular endothelial growth factor (VEGF) (Cat No DY293B), human monocyte chemoattractant protein-1 (MCP-1/CCL2) (Cat No DY279), human matrix metalloproteinase-9 (MMP-9) (Cat No DY911) and human angiopoietin-2 (Cat No DY623) were purchased from R&D Systems.

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: Enzyme-linked immunosorbent assay (ELISA) kits for human Hyal-1 (Cat No DY7358), human Hyaluronan (Cat No DY3614-05), and human syndecan-1 (Cat No DY2780), human vascular endothelial growth factor (VEGF) (Cat No DY293B), human monocyte chemoattractant protein-1 (MCP-1/CCL2) (Cat No DY279), human matrix metalloproteinase-9 (MMP-9) (Cat No DY911) and human angiopoietin-2 (Cat No DY623) were purchased from R&D Systems.

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