mouse monoclonal anti vegf antibody  (R&D Systems)

Journal: Investigative Ophthalmology & Visual Science

Article Title: Mathematical Models of Topically and Intravitreally Applied Ranibizumab

doi: 10.1167/iovs.66.11.45

Figure Lengend Snippet: Diagram of the experimental procedure. Porcine eyes were treated either topically or via intravitreal injection. The aqueous humor was collected using a syringe, while the vitreous humor was harvested after removal of the anterior segment. Ranibizumab and VEGF levels were detected using ELISA, and the optical density of the fluids was measured using a plate reader and then converted into a concentration using the respective standard curves. Figure created in https://BioRender.com/g17g888 .

Article Snippet: A total of 1 mg of humanized anti-VEGF monoclonal antibody fragments (ranibizumab) was purchased already constituted in PBS from MedChem Express (CAT: HY-P9951A-1mg; Princeton, NJ, USA) and stored at −80°C.

Techniques: Injection, Enzyme-linked Immunosorbent Assay, Concentration Assay

Journal: Investigative Ophthalmology & Visual Science

Article Title: Mathematical Models of Topically and Intravitreally Applied Ranibizumab

doi: 10.1167/iovs.66.11.45

Figure Lengend Snippet: Experimental results for treatment of ex vivo porcine eyes. Panels show ranibizumab (RBZ, first two columns ) and VEGF ( last two columns ) concentrations in the aqueous (Aq, columns 1 and 3 ) and vitreous (Vit, columns 2 and 4 ), at t = 20 min, 40 min, 1 h and 3.5 h. Top row : topical treatment (45 µL) with ranibizumab (1 mg mL −1 ) and CPP (100 mg mL −1 ). Ranibizumab is detected in significant quantities in the aqueous and appears to enter the vitreous, though it only has a significant effect in reducing aqueous VEGF levels, leaving vitreal VEGF levels unaffected. Middle row : intravitreal (invit.) treatment (45 µL) with ranibizumab (1 mg mL −1 ) and CPP (100 mg mL −1 ). As expected, vitreal ranibizumab levels are high, resulting in a significant reduction in vitreal VEGF levels. Bottom row : intravitreal treatment (45 µL) with ranibizumab only (1 mg mL −1 ). Interestingly, intravitreal treatment is more effective in reducing vitreal VEGF in the absence of CPPs. See Table 5 for numerical values of data points.

Article Snippet: A total of 1 mg of humanized anti-VEGF monoclonal antibody fragments (ranibizumab) was purchased already constituted in PBS from MedChem Express (CAT: HY-P9951A-1mg; Princeton, NJ, USA) and stored at −80°C.

Techniques: Ex Vivo

Journal: Investigative Ophthalmology & Visual Science

Article Title: Mathematical Models of Topically and Intravitreally Applied Ranibizumab

doi: 10.1167/iovs.66.11.45

Figure Lengend Snippet: Mathematical model fits to ex vivo porcine data. Panels show ranibizumab (R) concentrations in the aqueous (Aq, left column ) and vitreous (Vit, right column ), for topical drop administration with CPPs ( top row ) and for intravitreal injection without CPPs ( bottom row ). Top row : a single drop is applied at t = 0 h; simulations start at t = 0 h with r Tear (0) = r Dose = 2.07 × 10 4 pmol mL −1 , r Aq (0) = 0 pmol mL −1 and r Vit (0) = 0 pmol mL −1 ; fitting was performed for r Aq at t = 20 min, 40 min, and 3.5 h to the mean data points; Equations 3 – were solved with depleting tear ranibizumab concentration, constant tear volume, and in the absence of VEGF. Bottom row : a single injection is administered at t = 0 h; simulations start at t = 20 min (=1/3 h) with r Aq (1/3) = 4.73 × 10 −2 pmol mL −1 and r Vit (1/3) = 4.29 pmol mL −1 , equal to the mean data points at those times; fitting was performed for r Aq at t = 40 min, 1 h and 3.5 h to the mean data points; Equations 4 – were solved in the absence of VEGF. Reasonable fits are achieved in all cases except for vitreal ranibizumab with topical treatment. Model fit 1: β Tear-Aq, r = 5.93 × 10 −7 cm h −1 (this value is also used for model fit 2 in the topical case) and β Aq-Vit, r = 0.929 cm h −1 ; model fit 2: β Aq-Vit, r = 0.577 cm h −1 . All remaining parameters chosen as the default porcine values in Table 3 .

Article Snippet: A total of 1 mg of humanized anti-VEGF monoclonal antibody fragments (ranibizumab) was purchased already constituted in PBS from MedChem Express (CAT: HY-P9951A-1mg; Princeton, NJ, USA) and stored at −80°C.

Techniques: Ex Vivo, Injection, Concentration Assay

Journal: Investigative Ophthalmology & Visual Science

Article Title: Mathematical Models of Topically and Intravitreally Applied Ranibizumab

doi: 10.1167/iovs.66.11.45

Figure Lengend Snippet: Simulation results for treatment of an in vivo human eye, using a single mode of administration. Panels show VEGF (V) concentrations ( left column ) and ranibizumab (R) concentrations ( right column ) in the tear film (Tear), aqueous (Aq), and vitreous (Vit), as appropriate (insets show results over the full range of concentrations [ top right ], or with a logarithmic scale on the ordinate [ bottom row ]). Horizontal blue lines in the left column show untreated VEGF levels in the aqueous ( dotted ) and vitreous ( dash-dot ). Top row (topical drops): a single drop is applied at the start of hours 1 to 16 every day for the first 2 weeks; week 3 untreated. Middle row (drug-eluting contact lens): a series of four lenses are worn for 30 days at a time, starting on day 1, with a 1-day break between lenses; final 4 weeks untreated. Bottom row (intravitreal injections): administered at the start of weeks 1, 5, 9, and 13; simulation runs to 16 weeks. Drops suppress aqueous and vitreal VEGF levels to a fairly constant value during the period of administration; contact lenses suppress VEGF levels more strongly, with small transient increases in VEGF levels between lenses; injections reduce VEGF to by far the lowest levels, though VEGF returns to untreated values between injections. Equations 2 – were solved using the default human parameters in Table 3 .

Article Snippet: A total of 1 mg of humanized anti-VEGF monoclonal antibody fragments (ranibizumab) was purchased already constituted in PBS from MedChem Express (CAT: HY-P9951A-1mg; Princeton, NJ, USA) and stored at −80°C.

Techniques: In Vivo

Journal: Investigative Ophthalmology & Visual Science

Article Title: Mathematical Models of Topically and Intravitreally Applied Ranibizumab

doi: 10.1167/iovs.66.11.45

Figure Lengend Snippet: Simulation results for treatment of an in vivo human eye, using multiple modes of administration. Panels show VEGF (V) concentrations ( left column ) and ranibizumab (R) concentrations ( right column ) in the tear film (Tear), aqueous (Aq), and vitreous (Vit), as appropriate (insets show results over the full range of concentrations [ top right ], or with a logarithmic scale on the ordinate [ top left and bottom row ]). Horizontal blue lines in the left column show untreated VEGF levels in the aqueous ( dotted ) and vitreous ( dash-dot ). Top row (topical drops and intravitreal injections): a single drop is applied at the start of hours 1 to 16 every day for the first 16 weeks, while injections are administered at the start of weeks 1, 5, 9, and 13; simulation runs to 20 weeks, the final 4 untreated. Bottom row (drug-eluting contact lens and intravitreal injections): a series of four lenses are worn for 27 days at a time, starting on day 2, with a 1-day break between lenses, while injections are administered at the start of weeks 1, 5, 9, and 13 (on the days without contact lenses); simulation runs to 20 weeks, the final 4 untreated. Both drops and drug-eluting lenses suppress aqueous and vitreal VEGF levels between injections, preventing them from returning to untreated levels as in Figure 5 ( bottom row ). Equations 2 – were solved using the default human parameters in Table 3 .

Article Snippet: A total of 1 mg of humanized anti-VEGF monoclonal antibody fragments (ranibizumab) was purchased already constituted in PBS from MedChem Express (CAT: HY-P9951A-1mg; Princeton, NJ, USA) and stored at −80°C.

Techniques: In Vivo

Journal: Investigative Ophthalmology & Visual Science

Article Title: Mathematical Models of Topically and Intravitreally Applied Ranibizumab

doi: 10.1167/iovs.66.11.45

Figure Lengend Snippet: Effect of dosing regimen on vitreal VEGF and ranibizumab levels. Panels show variation in the maximum/mean/minimum vitreal VEGF concentration, v Vit ( top row ), and maximum/mean/minimum total ranibizumab concentration, r Tot, Vit = r Vit + u Vit + 2 w Vit ( bottom row ), in response to variation in dosing frequency or duration ( first–third columns : circles show the corresponding values for the dosing regimens plotted in Fig. 5 ). Top row : horizontal green dash-dot lines show the untreated vitreal VEGF level. First column (topical drops only): the number of drops per day, n drop , is varied across all integer values between 1 and 16 inclusive, where the first drop of each day is administered at 0 h, with subsequent drops being administered in time increments of 16/ n drop h. Simulated for t ∈ [0, 12] weeks, with plotted values calculated over model outputs from the interval t ∈ [9, 12] weeks. Second column (drug-eluting contact lenses only): lenses are worn for n day days at a time, starting on day 1, with a 1-day break between lenses, where n day is varied across all integer values between 1 and 30 inclusive. Simulated for t ∈ [0, 124] days (enough for four full treatment cycles of 31 days: 30 days on and 1 day off), with plotted values calculated over the final treatment cycle (of n day days on and 1 day off). Third column (intravitreal injections only): the time between injections is varied across all integer values between 1 and 8 weeks inclusive, with the first injection being administered at the start of week 1. Fourth column (topical drops and intravitreal injections): injections are administered as in the third column, with the addition of four topical drops per day (the realistic maximum frequency ) at 4, 8, 12, and 16 h. Fifth column (drug-eluting contact lenses and intravitreal injections): injections are administered as in the third column, with the addition of drug-eluting contact lenses worn for 6 days a week (the longest duration that can be used across all interinjection intervals while allowing for a 1-day break between lenses) between days 2 and 7 of each week. Third to fifth columns : simulated for t ∈ [0, 32] weeks (enough for four full treatment cycles lasting the maximum interval of 8 weeks), with plotted values calculated over model outputs from the final treatment cycle (with duration equal to the time between injections). Increasing drop or injection frequency, or increasing the time interval for which drug-eluting contact lenses are worn, increases the maximum, mean, and minimum vitreal total ranibizumab concentrations and decreases the maximum, mean, and minimum vitreal VEGF concentrations. Combining injections with drops or contact lenses visibly reduces the maximum and mean vitreal VEGF concentrations compared to injections alone, while the effect on the minimum vitreal VEGF concentration and the maximum, mean, and minimum vitreal total ranibizumab concentrations is more subtle. Equations 2 – were solved using the default human parameters in Table 3 .

Article Snippet: A total of 1 mg of humanized anti-VEGF monoclonal antibody fragments (ranibizumab) was purchased already constituted in PBS from MedChem Express (CAT: HY-P9951A-1mg; Princeton, NJ, USA) and stored at −80°C.

Techniques: Concentration Assay, Injection

Journal: Investigative Ophthalmology & Visual Science

Article Title: Mathematical Models of Topically and Intravitreally Applied Ranibizumab

doi: 10.1167/iovs.66.11.45

Figure Lengend Snippet: Local sensitivity analysis. Panels show sensitivity of the maximum/mean/minimum vitreal VEGF (V) concentration ( left column ) and total vitreal ranibizumab (R Tot = R + VR + 2RVR) concentration ( right column ) to variation in model parameters over biologically realistic ranges (insets show full range of sensitivity values). Equations 2 – were solved for t ∈ [0, 12] weeks. Top row : topical drops, applied on the hour, every hour. Middle row : drug-eluting contact lens, worn continuously. Bottom row : intravitreal injections, administered at the start of weeks 1, 5, and 9. Parameters were varied individually, across 101 values uniformly distributed over the ranges given in Table 4 , with the remaining parameters being held at their default values given in Table 4 . For each parameter set, the maximum/mean/minimum vitreal values of V and R Tot were calculated over model outputs from the interval t ∈ [9, 12] weeks (see Supplementary Figs. S2 – ). Each sensitivity factor was then calculated as the maximum value obtained by the maximum/mean/minimum value of V or R Tot over the 101 values across which the parameter was varied, divided by the corresponding minimum value over that range. The dashed red horizontal line demarcates the sensitivity threshold (=1.5), above which sensitivity is considered significant. The model demonstrates sensitivity to the following parameters—drops: ( V Tear Norm , A Aq - Vit , k + , δ Aq , r , δ Vit , v , δ Vit , r , ϕ Vit , v , ψ Tear , ψ Aq ) ; contact lens: ( A Tear-Aq , k + , δ Aq, r , δ Vit, v , δ Vit, r , ϕ Vit, v , ψ Aq ), injections: ( V Vit , A Aq-Vit , k + , δ Aq, r , δ Vit, v , δ Vit, r , δ Vit, u , β Aq-Vit, r , ϕ Vit, v , ψ Aq ).

Article Snippet: A total of 1 mg of humanized anti-VEGF monoclonal antibody fragments (ranibizumab) was purchased already constituted in PBS from MedChem Express (CAT: HY-P9951A-1mg; Princeton, NJ, USA) and stored at −80°C.

Techniques: Concentration Assay

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: Acta neuropathologica communications

Article Title: Development of CAR-T cell therapy for NF1/SWN-related nerve sheath tumor treatment.

doi: 10.1186/s40478-025-01965-6

Figure Lengend Snippet: Fig. 1 Expression analysis of 6 antigens in 85 benign NF1/SWN-related nerve sheath tumor samples by IHC staining. A Expression intensity of HER1, HER2, VEGFR2, B7H3, CD171 and EGFRvIII in 25 NF1 and 60 SWN-NOS tumor samples. B Typical images of HER1, HER2, VEGFR2, B7H3, CD171 and EGFRvIII expression in 3 NF1 and 6 SWN-NOS tumor samples respectively. Scale bar = 100 μm. NF1, type 1 neurofibromatosis; SWN-NOS, schwannomatosis-not otherwise specified

Article Snippet: Immunohistochemistry (IHC) was performed at ZSGBBIO (Beijing, China) on 5-μm FFPE human tissue and spheroid sections using the following unconjugated antibodies: rabbit anti-human HER1 (Abcam, ab52894), rabbit anti-human HER2 (ZSGB-BIO, ZA-0023), rabbit anti-human VEGFR2 (CST, 9698S), rabbit anti-human B7H3 (CST, #14058), rabbit anti-human CD171 (Abcam, ab208155), rabbit anti-human EGFRvIII (ZSGB-BIO, ZA-0643), rabbit anti-human TGFβ1 (Abcam, ab215715), rabbit anti-human PDL1 (Genetex, GTX104763) and rabbit anti-human S100 (ZSGB-BIO, ZA-0225).

Techniques: Expressing, Immunohistochemistry

Journal: Biochemistry and Biophysics Reports

Article Title: In vitro analysis of VEGF-mediated endothelial permeability and the potential therapeutic role of Anti-VEGF in severe dengue

doi: 10.1016/j.bbrep.2024.101814

Figure Lengend Snippet: Fig. 1. In vitro permeability assay. (A) Brightfield image of human pulmonary microvascular endothelial cells (HPMECs) during culture before seeding for permeability assay. Scale bar = 200 μm. (B) Graph of fluorescein isothiocyanate (FITC)-dextran fluorescence signal over time (n = 3). Asterisks indicate a significant increase in signal from VEGF-treated cells compared to untreated cells at *p < 0.01. Hashtags indicate a significant reduction in signal from VEGF/anti-VEGF-treated cells compared to VEGF-treated cells at #p < 0.01. A higher FITC-dextran signal represents increased movement over the endothelial layer and greater endothelial permeability. VEGF-treated cells showed higher endothelial permeability compared to untreated cells and VEGF/anti-VEGF-treated cells. (C) Immunostaining images of untreated and treated cells (incubated for 180 min) stained with PECAM-1 (n = 3). The formation of gaps between cells allowed FITC-dextran to cross the endothelial cell monolayer. Anti-VEGF treatment reduced the number of gaps formed between cells. Scale bar = 25 μm.

Article Snippet: The VEGF used in this study was a recombinant VEGF protein (Catalogue # GF615, Merck, Darmstadt, Germany), whereas the anti-VEGF used was a human VEGF monoclonal antibody (Catalogue # MAB293, R&D Systems, MN, USA).

Techniques: In Vitro, Permeability, Fluorescence, Immunostaining, Incubation, Staining

Journal: Biochemistry and Biophysics Reports

Article Title: In vitro analysis of VEGF-mediated endothelial permeability and the potential therapeutic role of Anti-VEGF in severe dengue

doi: 10.1016/j.bbrep.2024.101814

Figure Lengend Snippet: Fig. 2. Volcano plot of differentially expressed genes (DEGs) in VEGF-treated cells (left) and VEGF/anti-VEGF-treated cells (right). The plot showed top 10 upre gulated and downregulated genes. Red represents upregulated genes, blue represents downregulated genes, and black represents genes with no significant change in fold change. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: The VEGF used in this study was a recombinant VEGF protein (Catalogue # GF615, Merck, Darmstadt, Germany), whereas the anti-VEGF used was a human VEGF monoclonal antibody (Catalogue # MAB293, R&D Systems, MN, USA).

Techniques:

Journal: Biochemistry and Biophysics Reports

Article Title: In vitro analysis of VEGF-mediated endothelial permeability and the potential therapeutic role of Anti-VEGF in severe dengue

doi: 10.1016/j.bbrep.2024.101814

Figure Lengend Snippet: Fig. 5. Protein-protein interaction (PPI) network of differentially expressed genes. A PPI network identified from DEGs of VEGF-treated endothelial cells using STRING. This PPI network was used to identify hub genes using Cytoscape, interaction score >0.4. Top 10 hub genes for VEGF-treated endothelial cells identified by CytoHubba, plugin in Cytoscape, using MCC algorithm. All the genes identified were upregulated genes in VEGF-treated cells. Red represents genes with high MCC score and yellow represents genes with low MCC score. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: The VEGF used in this study was a recombinant VEGF protein (Catalogue # GF615, Merck, Darmstadt, Germany), whereas the anti-VEGF used was a human VEGF monoclonal antibody (Catalogue # MAB293, R&D Systems, MN, USA).

Techniques:

Journal: Frontiers in Cell and Developmental Biology

Article Title: Syndecan-1 as a predictor of vulnerable atherosclerotic plaques

doi: 10.3389/fcell.2024.1415788

Figure Lengend Snippet: Analysis of destabilizing components in atherosclerotic plaque. (A) The expression of lipid deposition in aortic root plaques was detected by oil red O with a scale bar of 100 μm, immunofluorescence detection of CD68 expression (macrophage marker) and VEGF in aortic root plaques with a scale bar of 20 μm, and IHC detection of MMP-9 expression in aortic root plaques with a scale bar of 100 μm. (B–E) Statistical chart for quantitative analysis of oil red o staining, CD68, VEGF and MMP-9 in the aortic root plaques (n = 6, one-way ANOVA). (F) Immunofluorescence detection of BODIPY and VEGF expression in LCCA plaques with a scale bar of 100 μm. IHC detection of CD68 and MMP-9 expression in LLCCA plaques with a scale bar of 100 μm. (G–J) Statistical chart for quantitative analysis of BODIPY staining, VEGF, CD68 and MMP-9 in the LCCA plaques (n = 6, unpaired t -test). * P < 0.05, ** P < 0.01, *** P < 0.001, compared with CD group; # P < 0.05, ## P < 0.01, ### P < 0.001, compared with CD + PLCA group; && P < 0.01, &&& P < 0.001, compared to HFD group.

Article Snippet: After blocking, sections were incubated with a rabbit anti-human CD68 (1:100, bs-4819R, BIOSS, China), α-SMA (1:100, A1011, Abclonal, China) and mouse anti-human VEGF (1:100, TA500289, Origene, United States of America) antibodies overnight at 4°C.

Techniques: Expressing, Immunofluorescence, Marker, Staining

Journal: Frontiers in Cell and Developmental Biology

Article Title: Syndecan-1 as a predictor of vulnerable atherosclerotic plaques

doi: 10.3389/fcell.2024.1415788

Figure Lengend Snippet: Summary of the plaque components in HFD and PLCA apoE −/− mice.

Article Snippet: After blocking, sections were incubated with a rabbit anti-human CD68 (1:100, bs-4819R, BIOSS, China), α-SMA (1:100, A1011, Abclonal, China) and mouse anti-human VEGF (1:100, TA500289, Origene, United States of America) antibodies overnight at 4°C.

Techniques:

Journal: Frontiers in Cell and Developmental Biology

Article Title: Syndecan-1 as a predictor of vulnerable atherosclerotic plaques

doi: 10.3389/fcell.2024.1415788

Figure Lengend Snippet: Serum SDC1 is an independent predictor of high-risk vulnerable plaque. (A) Serum SDC1; (B) S1P; (C) VEGF-A was detected using ELISA after 16 weeks of modeling, n = 6, one-way ANOVA. (D) ROC curve. (E) Multifactor binary logistic regression analysis was performed on serum SDC1, S1P and VEGF-A. (F) Receiver operating characteristic (ROC) curve analysis was used to evaluate the diagnostic accuracy of the variables. The Jorden index, calculated as sensitivity minus (1 - specificity), was used to determine the optimal cut-off point on the ROC curve. The area under the ROC curve (AUC) was utilized to assess the clinical prediction value of SDC1. Notes: AUC, area under the curve; PV+, positive predictive value; PV-, negative predictive value. * P < 0.05, * P < 0.01, *** P < 0.001, compared with CD group; # P < 0.05, compared with CD + PLCA group; && P < 0.01, compared to HFD group.

Article Snippet: After blocking, sections were incubated with a rabbit anti-human CD68 (1:100, bs-4819R, BIOSS, China), α-SMA (1:100, A1011, Abclonal, China) and mouse anti-human VEGF (1:100, TA500289, Origene, United States of America) antibodies overnight at 4°C.

Techniques: Enzyme-linked Immunosorbent Assay, Diagnostic Assay

Journal: Acta biomaterialia

Article Title: In vitro throughput screening of anticancer drugs using patient-derived cell lines cultured on vascularized three-dimensional stromal tissues.

doi: 10.1016/j.actbio.2024.05.037

Figure Lengend Snippet: Fig. 4. Evaluation of drug efficacy using cancer cell line models generated by the CAViTs method Comparison of drug efficacy evaluations using cell line models generated using the CAViTs method, the 2D method, and spheroid culture method in 96-well plates (black: L-OHP only, gray: L-OHP+VEGF monoclonal antibody [mAb], HT29) (a). Comparison of drug efficacy evaluation results with and without a vascular network (black: 5-FU only, gray: 5-FU+VEGF mAb, HCT116) (b). Comparison of efficacy evaluation results for HUVEC content (black: L-OHP only, gray: L-OHP+VEGF mAb, HCT116) (c). **P<0.05, n.s.: not significant.

Article Snippet: Antibodies included anti–vascular endothelial growth factor (VEGF) monoclonal antibody (MAB293, R&D Systems), EpCAM monoclonal antibody (#2929S, Cell Signaling Technology Corp.), cytokeratin 7 monoclonal antibody (ab181598, Abcam PLC.), Pan Cytokeratin Monoclonal Antibody (MA5-13156, Invitrogen.), carcinoembryonic antigen (CEA)/CD66e monoclonal antibody (#2383, Cell Signaling Technology Corp.), Non-phospho (Active) β-Catenin(#8814, Cell Signaling Technology Corp.), CK8/18 monoclonal antibody (M3652, Agilent Corp.), platelet endothelial cell adhesion molecule (CD31) monoclonal antibody (M0823, Agilent Corp.), and anti–hepatocyte growth factor (HGF) antibody (MAB294, R&D Systems Corp.).

Techniques: Generated, Comparison

Journal: Acta biomaterialia

Article Title: In vitro throughput screening of anticancer drugs using patient-derived cell lines cultured on vascularized three-dimensional stromal tissues.

doi: 10.1016/j.actbio.2024.05.037

Figure Lengend Snippet: Fig. 6. Construction of PDC-stromal models using the CAViTs method and evaluation of drug efficacy IHC images of colorectal JC-143-1–stromal models generated using the CAViTs method (brown: CEA, gray: CD31, white inset: cancer invasion into blood vessel) (a) and enlarged image of the cancer invasion area (b). IHC images of colorectal JC-143-1–stromal models (brown: CEA, arrow: glandular duct structure) (c). HE image (dashed white area: nucleus of cancer cells on the stromal side) (d). Results of drug efficacy evaluation using colorectal JC-143-1–stromal models (black: 5-FU only, gray: 5-FU+VEGF antibody) (e). Immunofluorescence images of colorectal JC-143-1–stromal models with and without Bmab (green: EpCAM, red: CD31) (f, g). Comparison of the results of efficacy evaluations of a two-drug combination (5-FU+L-OHP) between the CAViTs method and 2D culture method (black: CAViTs, gray: 2D) (h). Comparison of the results of efficacy evaluation of a three-drug combination (5-FU+L-OHP+Cmab) (black: CAViTs, gray: 2D) (i). *:0.05

Article Snippet: Antibodies included anti–vascular endothelial growth factor (VEGF) monoclonal antibody (MAB293, R&D Systems), EpCAM monoclonal antibody (#2929S, Cell Signaling Technology Corp.), cytokeratin 7 monoclonal antibody (ab181598, Abcam PLC.), Pan Cytokeratin Monoclonal Antibody (MA5-13156, Invitrogen.), carcinoembryonic antigen (CEA)/CD66e monoclonal antibody (#2383, Cell Signaling Technology Corp.), Non-phospho (Active) β-Catenin(#8814, Cell Signaling Technology Corp.), CK8/18 monoclonal antibody (M3652, Agilent Corp.), platelet endothelial cell adhesion molecule (CD31) monoclonal antibody (M0823, Agilent Corp.), and anti–hepatocyte growth factor (HGF) antibody (MAB294, R&D Systems Corp.).

Techniques: Generated, Immunofluorescence, Comparison