recombinant mouse vcam1 fc  (R&D Systems)

Journal: Journal of Inflammation Research

Article Title: Mechanisms of Yiqi Huoxue Granule in Atherosclerosis Treatment: Insights from UPLC-Q-Exactive Orbitrap-MS Analysis, Network Pharmacology, Molecular Docking, and Experimental Verification

doi: 10.2147/JIR.S566368

Figure Lengend Snippet: Effects of YQHX on serum inflammatory cytokines and macrophage infiltration within atherosclerotic plaques in ApoE –/– mice. ( A–D ) Concentrations of TNF-α, IL-6, IL-1β, and sVCAM-1 in serum (n = 6). ( E ) Immunohistochemistry staining of F4/80 in plaques (n = 3). Data are expressed as means ± SD. ### P < 0.001 versus control, * P < 0.05 versus model, ** P < 0.01 versus model, *** P < 0.001 versus model.

Article Snippet: According to the manufacturer’s protocols for TNF-α (Cat. #EK282, Lianke), IL-6 (Cat. #EK206, Lianke), interleukin-1 beta (IL-1β; Cat. #EK201B, Lianke), and soluble VCAM-1 (sVCAM-1) (Cat. #EK290, Lianke) ELISA kits, mouse serum samples were introduced into microplate wells and incubated with a diluted primary antibody solution.

Techniques: Immunohistochemistry, Staining, Control

Journal: bioRxiv

Article Title: A systematic cross-modal approach identifies astrocytic VCAM1 as a regulator of hippocampal synapse development

doi: 10.64898/2026.03.17.712320

Figure Lengend Snippet: (A) Schematic of the workflow used to assess the effect of constitutive CRISPR/Cas9-mediated KO of Vcam1 or Gpr37l1 on hippocampal synapse development. SO: stratum oriens, SR: stratum radiatum, SLM: stratum lacunosum-moleculare, SL: stratum lucidum, DG: dentate gyrus, ML: molecular layer. Created with BioRender.com. (B) Quality control of Vcam1 KO by immunohistochemistry for VCAM1 and DAPI. Insets show CA1 region of both hemispheres. (C) Quality control of Gpr37l1 KO by immunohistochemistry for GPR37L1 and DAPI. Insets show CA1 region of both hemispheres. (D) Heatmap showing the log2 fold change in synaptic metrics comparing LacZ-gRNA and VCAM1-gRNA hemispheres across hippocampal laminae. For each synaptic metric and lamina, 3 FOVs (sections) per brain from 4 brains were analyzed. Log2-transformed metrics were analyzed using a linear mixed-effects model with gRNA, hippocampal lamina and their interaction as fixed effects to test gene- and lamina-specific effects, and brain as a random intercept to account for inter-brain variability. Asterisks denote significant differences after false discovery rate (FDR) correction for each metric across hippocampal laminae (FDR < 0.05). (E) Heatmap showing the log2 fold change in synaptic metrics comparing LacZ-gRNA and GPR37L1-gRNA hemispheres. Same sampling, plotting and analyses as in (G). (F) Representative processed Airyscan images of VGLUT1 and PSD95 in CA1 SR from LacZ-gRNA- and VCAM1-gRNA-injected hemispheres, with quantification of local peak max and presynapse MFI normalized to the LacZ-gRNA hemisphere. Each small data point represents a single FOV; larger linked data points represent hemisphere means. Statistical significance was assessed using a linear mixed effects model as in (G), followed by Tukey’s post hoc tests and FDR corrected significance. (G) Representative processed Airyscan images of VGLUT1 and PSD95 in CA3 SR from LacZ-gRNA- and VCAM1-gRNA-injected hemispheres, with quantification of local peak max and presynapse MFI normalized to the LacZ-gRNA hemisphere. Same sampling, plotting and analyses as in (F). (H) Schematic of the workflow used to assess the effect of VCAM1-Fc protein treatment on hippocampal synapses in vitro . Created with BioRender.com. (I) Representative images of hippocampal neurons treated with recombinant protein from DIV8 to DIV14. Fc or VCAM1-Fc (2 µg/mL) was added at DIV8 and replenished at DIV11. Neurons were fixed at DIV14 and stained for VGLUT1 (cyan), PSD95 (magenta), and MAP2 (yellow). Insets show the merge followed by the isolated VGLUT1-PSD95 signal. (J) Quantification of excitatory synaptic puncta density along MAP2+ dendrites. VGLUT1+PSD95+ colocalized puncta, as well as VGLUT1+ and PSD95+ puncta separately, were measured per 100 μm dendrite and normalized to the Fc condition. Each FOV contained two neuronal somas for standardized sampling; 9-11 FOVs per condition were analyzed in N = 4 independent experiments. Each data point represents one experiment is color-shaded by experiment. A linear mixed-effects model on log2 transformed data was used to account for inter-experimental variability. Post-hoc pairwise comparisons were conducted using Tukey’s HSD test. Individual FOV data points are shown in .

Article Snippet: Upon arrival, lyophilized recombinant mouse VCAM1-Fc (R&D Systems, 643-VM) was briefly centrifuged and reconstituted in sterile Dulbecco’s phosphate-buffered saline (DPBS).

Techniques: CRISPR, Control, Immunohistochemistry, Transformation Assay, Sampling, Injection, In Vitro, Recombinant, Staining, Isolation

Journal: bioRxiv

Article Title: A systematic cross-modal approach identifies astrocytic VCAM1 as a regulator of hippocampal synapse development

doi: 10.64898/2026.03.17.712320

Figure Lengend Snippet: (A) Western blot quality control of the recombinant proteins Fc and VCAM1-Fc used to treat hippocampal neurons, visualized using a total protein stain. (B) Representative images of hippocampal neurons treated with recombinant protein from DIV8 to DIV14. Fc or VCAM1–Fc (2 µg/mL) was added at DIV8 and replenished at DIV11. Neurons were fixed at DIV14 and stained for VGAT (cyan), GEPH (magenta), and MAP2 (yellow). Insets show the merge followed by the isolated VGAT-GEPH signal. (C) Quantification of inhibitory synaptic puncta density along MAP2+ dendrites. VGAT+GEPH+ colocalized puncta, as well as VGAT+ and GEPH+ puncta separately, were measured per 100 μm dendrite and normalized to Fc. Same sampling, plotting and analyses as in . (D) Plots from showing distribution of the analyzed FOVs. Each dot represents a single FOV and is color-shaded according to experiment. (E) Plots from (C) showing distribution of the analyzed FOVs. Each dot represents a single FOV and is color-shaded according to experiment. (F) Quantification of excitatory synaptic puncta size of VGLUT1+PSD95+ colocalized puncta, as well as VGLUT1+ and PSD95+ puncta separately. Each FOV contained two neuronal somas for standardized sampling; 9-11 FOVs per condition were analyzed in N = 4 independent experiments. Each data point represents one experiment (above panel), and one FOV (below panel), color-shaded by experiment. A linear mixed-effects model was used to account for multiple FOVs per experiment. (G) Quantification of inhibitory synaptic puncta size of VGAT+GEPH+ colocalized puncta, as well as VGAT+ and GEPH+ puncta separately. Same sampling, plotting and analyses as in (F).

Article Snippet: Upon arrival, lyophilized recombinant mouse VCAM1-Fc (R&D Systems, 643-VM) was briefly centrifuged and reconstituted in sterile Dulbecco’s phosphate-buffered saline (DPBS).

Techniques: Western Blot, Control, Recombinant, Staining, Isolation, Sampling

Journal: Advanced Healthcare Materials

Article Title: Simvastatin Restores Uteroplacental Hemodynamics and Trophoblast Function in Obstetric Antiphospholipid Syndrome in a Placenta‐on‐a‐Chip Model

doi: 10.1002/adhm.202504027

Figure Lengend Snippet: Simvastatin Ameliorates Decidual Vessel Damage and Improves Placental Perfusion in Obstetric Antiphospholipid Syndrome (OAPS) Mice. (A) Experimental design of the OAPS mouse model and simvastatin treatment protocol. (B) Uterine morphology at E14.5; arrows indicate sites of fetal resorption. (C) Histopathological assessment of placental inflammation and structural abnormalities in the decidua basalis (D), junctional zone (JZ), and labyrinth (L); arrows in the hematoxylin and eosin ( H&E )‐stained decidual region highlight diseased vessels and fibrinoid necrosis. Scale bar, 200 µm. (D) Quantification of fetal resorption frequency (FRF) at E14.5 ( n = 6, one‐way ANOVA). (E) Statistical analysis of placental weight among the three groups at E14.5 ( n = 7, one‐way ANOVA). (F,G) Representative images and quantification of interleukin ( IL )‐1β and IL‐6 immunofluorescent staining in the decidua ( n = 5, two‐way ANOVA for IL‐6 vs. IL‐1β) (Red: IL‐6; Green: IL‐1β; Blue: DAPI). Scale bar, 100 µm. (H,I) Representative images and quantification of vascular cell adhesion molecule 1 (VCAM‐1) and intercellular adhesion molecule 1 (ICAM‐1) immunofluorescent staining in the decidua ( n = 5, two‐way ANOVA for VCAM‐1 vs. ICAM‐1) (Red: VCAM‐1; Green: ICAM‐1; Blue: DAPI). Scale bar, 100 µm. J, L) Representative low‐ and high‐magnification images of endothelial nitric oxide synthase ( eNOS ) immunohistochemical staining in decidual vessels with corresponding mean density quantification ( n = 5, one‐way ANOVA). Scale bar, 100 µm. (K,M) Representative images of placental perfusion fluorescence intensity and corresponding quantification ( n = 5, one‐way ANOVA). Scale bar, 100 µm. Data are presented as mean ± standard deviation (SD). Statistical significance: ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Primary antibodies: KLF2 (512217, CST); eNOS (95867, CST); VCAM‐1 (TA502391S, ORIGENE); ICAM‐1 (67836, CST); E‐Selectin (ER1908‐35, HuanBiotechnology); MMP‐2 (TA806846S); Beta‐Actin (66009‐1‐Ig, Proteintech); Tubulin (10094‐1‐AP, Proteintech).

Techniques: Staining, Immunohistochemical staining, Fluorescence, Standard Deviation

Journal: Advanced Healthcare Materials

Article Title: Simvastatin Restores Uteroplacental Hemodynamics and Trophoblast Function in Obstetric Antiphospholipid Syndrome in a Placenta‐on‐a‐Chip Model

doi: 10.1002/adhm.202504027

Figure Lengend Snippet: Simvastatin Ameliorates aPL‐Induced Endothelial Dysfunction via the KLF2/eNOS Signaling Axis. (A) Assessment of HUVEC viability following simvastatin treatment ( n = 3, one‐way ANOVA). (B) Western blot analysis of adhesion molecules (VCAM‐1, ICAM‐1, E‐selectin). (C,L) Flow cytometric analysis of intracellular reactive oxygen species (ROS) levels. D–F) Densitometric quantification of VCAM‐1, ICAM‐1, and E‐selectin protein expression ( n = 3, one‐way ANOVA). (G–I) Relative mRNA expression of VCAM‐1 , ICAM‐1 , and E‐selectin determined by quantitative reverse transcription PCR (qRT‐PCR) ( n = 6, one‐way ANOVA). J, M) Representative DAF‐FM DA staining for nitric oxide (NO) and corresponding quantification ( n = 6, one‐way ANOVA). Scale bar, 100 µm. (K,N) Transwell invasion assay of HTR‐8/SVneo cells cocultured with HUVECs ( n = 3, one‐way ANOVA). Scale bar, 100 µm. (O,P) ELISA quantification of IL‐6 and IL‐1β secretion. (Q) Western blot analysis of Kruppel‐like factor 2 (KLF2) and eNOS following KLF2 small interfering RNA (siRNA) transfection. (R,S) Densitometric quantification of KLF2 and eNOS protein levels in siNC ‐ vs. siKLF2 ‐transfected HUVECs ( n = 3, one‐way ANOVA). (T,U) qRT‐PCR quantification of KLF2 and eNOS mRNA expression ( n = 6, one‐way ANOVA). Data are presented as mean ± SEM. Statistical significance: ns P ≥ 0.05, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Primary antibodies: KLF2 (512217, CST); eNOS (95867, CST); VCAM‐1 (TA502391S, ORIGENE); ICAM‐1 (67836, CST); E‐Selectin (ER1908‐35, HuanBiotechnology); MMP‐2 (TA806846S); Beta‐Actin (66009‐1‐Ig, Proteintech); Tubulin (10094‐1‐AP, Proteintech).

Techniques: Western Blot, Expressing, Reverse Transcription, Quantitative RT-PCR, Staining, Transwell Invasion Assay, Enzyme-linked Immunosorbent Assay, Small Interfering RNA, Transfection

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 microvascular endothelial cells (HRMECs) were left untreated or stimulated with tumor necrosis factor-α (TNF-α) (5 ng/mL) for 24 h with or without apigenin (10 µg/mL). Protein expression of intercellular adhesion molecule-1 (ICAM-1) (A) and vascular cell adhesion molecule-1 (VCAM-1) (B) was determined by Western blot analysis. Adhesion of fluorescently labeled THP-1 monocytic cells to HRMECs monolayer was quantified (C) . 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 values obtained from cells treated with TNF-α (RFU = relative fluorescence unit).

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, Western Blot, Labeling, Standard Deviation, Fluorescence

Journal: Frontiers in Toxicology

Article Title: Effect of the cysteine prodrug L-2-oxothiazolidine-4-carboxylic acid on in vivo platelet activation, oxidative stress, and procoagulant responses induced by waterpipe smoke exposure in mice

doi: 10.3389/ftox.2026.1696863

Figure Lengend Snippet: Plasma P-selectin (A) , E-selectin (B) , vascular cell adhesion molecule 1 [VCAM-1, (C) ] and intercellular adhesion molecule 1 [ICAM-1, (D) ] at the end of at the end of 1-month exposure period to air (control) or waterpipe smoke (WPS), with or without L-2-oxothiazolidine-4-carboxylic acid (OTC) treatment. Data are mean ± SEM (n = 8).

Article Snippet: The plasma concentrations of CRP (DY1829), PF4 (DY595), PAI-1 (DY3828), P-selectin (DY737), E-selectin, ICAM-1 (DY796), and VCAM-1 (DY643) were measured using commercially available ELISA kits from R&D Systems (DuoSet; Minneapolis, MN, United States).

Techniques: Clinical Proteomics, Control