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  • 93
    RayBiotech vcam 1
    Vcam 1, supplied by RayBiotech, used in various techniques. Bioz Stars score: 93/100, based on 36 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/vcam 1/product/RayBiotech
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    91
    ABclonal vcam 1
    Effects of ImKTx88 on ICAM-1 and <t>VCAM-1</t> expression. a Representative immunohistochemistry of ICAM-1 and VCAM-1 expression in the cerebellar white matter of EAE-induced and ImKTx88 treated rats. The scale bar represents 100 µm. Staining was quantified by mean of the integrated optical density (IOD) (5 random images per section and n = 3). b A representative western blot of cerebellum homogenates and quantification analysis revealed that the levels of ICAM-1 and VCAM-1 were significantly increased in EAE-induced rats, and ImKTx88 could decrease the levels of ICAM-1 and VCAM-1 (n = 5–12). GAPDH was used as an internal loading control. Data represent the mean ± SEM, * P
    Vcam 1, supplied by ABclonal, used in various techniques. Bioz Stars score: 91/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Cell Signaling Technology Inc vcam1
    TNF-induced <t>VCAM1</t> expression was decreased in TXNIP-deficient mouse aorta. After aortae from HcB-19 and control C3H mice were treated with TNF (15 ng/ml, 6 hours), vessel protein was harvested. Expression of VCAM1 and TXNIP was determined by immunoblotting
    Vcam1, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 14 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    Cell Signaling Technology Inc anti vcam1
    Effect of EPC Exosomal miR-126-3p and 5p on LPS-Induced HMVEC Target Expression and CLP-Induced Mortality Protein levels of <t>VCAM1</t> (A) and HMGB1 (B) in HMVECs were measured by western blot. α-Tubulin served as an internal control. * p
    Anti Vcam1, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 95/100, based on 26 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Thermo Fisher vcam1
    Gene ontology and localization of EC subtypes elucidates known functions and confirms divergence of clusters 2 and 4. ( A ) Circos plot shows gene expression and gene ontology (GO) overlap between distinct clusters of pulmonary ECs. A purple line connecting two clusters indicates expression of the same gene in both clusters, while a blue line connecting two clusters indicates expression of different genes found within the same GO category in each cluster. ( B ) GO biological process enrichment performed for each cluster and displayed in heatmap format demonstrates expression of genes associated with different biological processes in each cluster, including some overlap between clusters. Cluster 2 (green box) shares enrichment of some processes related to angiogenesis and blood vessel development with cluster 0 (blue box) but is distinct in its enrichment of genes related to vasculogenesis. ( C ) In-depth analysis of gene expression in cluster four indicates that this cluster likely represents macrovascular ECs (maECs) with high expression of Vwf and <t>Vcam1</t> . IHC indicates that these proteins localize mainly to the large vessel endothelium. White arrowhead demonstrates Vcam1 located in nearby mesenchymal cells. ( D ) Clusters 0, 1, and three represent a heterogeneous population of microvascular ECs (miECs) with high expression of Gpihbp1 and Plvap . IHC indicates that these proteins localize to the alveolar capillary plexus endothelium. Yellow arrowhead demonstrates Plvap present in the large vessel endothelium. ( E ) Cluster two represents an as-yet uncharacterized population of ECs that localize to the alveolar region and express surface marker Cd34 at a higher level from that in other ECs. These cells also express high levels of Car4 . CD34 and Car4 proteins localize to the alveolar space, indicating a similar spatial distribution of cells in cluster two to that of miECs. v, vessel; a, alveolar space; scale bars in ( C )-( E ), 20 microns.
    Vcam1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 91/100, based on 81 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Becton Dickinson vcam 1
    Total and CD11b lo macrophage subsets are greatly reduced before and during E-stress response in Spi-C–deficient and <t>VCAM-1–deficient</t> mice. Numbers of CD11b lo (lighter shades) and CD11b hi (darker shades) macrophages, as well as VCAM-1 expression
    Vcam 1, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 92/100, based on 688 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Santa Cruz Biotechnology vcam 1
    Immunohistochemical staining for F4/80-positive monocytes-derived macrophages and adhesion molecule <t>VCAM-1</t> in aortic cross-sections Representative photomicrographs of immunohistochemical staining for F4/80-positive monocytes-derived macrophages (Fig. 6A) and VCAM-1 (Fig. 6B). Quantitative analysis of F4/80 (Fig. 6C) and VCAM-1 (Fig. 6D). Arrows indicate typical positive stained regions and original magnification is 40×. T, TNF-α; T +G, TNF-α + genistein; Data are expressed as mean ± SEM, n=5, *, p
    Vcam 1, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 92/100, based on 2249 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    RayBiotech mouse vcam1 elisa kit
    Immunohistochemical staining for F4/80-positive monocytes-derived macrophages and adhesion molecule <t>VCAM-1</t> in aortic cross-sections Representative photomicrographs of immunohistochemical staining for F4/80-positive monocytes-derived macrophages (Fig. 6A) and VCAM-1 (Fig. 6B). Quantitative analysis of F4/80 (Fig. 6C) and VCAM-1 (Fig. 6D). Arrows indicate typical positive stained regions and original magnification is 40×. T, TNF-α; T +G, TNF-α + genistein; Data are expressed as mean ± SEM, n=5, *, p
    Mouse Vcam1 Elisa Kit, supplied by RayBiotech, used in various techniques. Bioz Stars score: 93/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Bender MedSystems vcam 1
    <t>VCAM-1</t> changes for massage group and control group. * P
    Vcam 1, supplied by Bender MedSystems, used in various techniques. Bioz Stars score: 92/100, based on 63 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Biosensis vcam 1
    Time course of ICAM-1 and <t>VCAM-1</t> expression in the rat AVM model. Xenolight-750 probe binding was examined over a period of 84 days for ICAM-1 (A) and VCAM-1 (B) in control and irradiated (15 Gy) animals (n = 7 per group). Raw MFI (mean fluorescence intensity) was normalized to day 1 in matched animals to reduce inter-animal variation. No significant differences were detected at any time between irradiated and control AVMs. Ex vivo image analysis of Xenolight 750-ICAM-1 (C) and Xenolight 750-VCAM-1 (D) probe binding in excised tissue: common carotid artery (CCA), external jugular vein (EJV) (n = 7).
    Vcam 1, supplied by Biosensis, used in various techniques. Bioz Stars score: 91/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Targeson vcam 1
    Summary of longitudinal changes in molecular US imaging measurements using either a P-selectin ( a ) or <t>VCAM-1</t> ( b ) targeted MB contrast agent. Bar graphs detail differences between ischemic and control kidney data. (* p
    Vcam 1, supplied by Targeson, used in various techniques. Bioz Stars score: 92/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    rdi research diagnostics vcam 1
    EC–substratum interaction influences NF-κB-dependent expression of adhesion molecules. Real-time PCR ΔΔCt of (A) <t>VCAM-1</t> and (B) ICAM in MEEC (opened histogram) and EC-TCPS (closed histogram) stimulated for 6 h with 5 ng/ml TNF-α. Flow cytometry analysis of (C) VCAM and (D) ICAM expression on embedded EC (solid line), TCPS (dashed line), and TCPS with SC514 100 µM (dotted line) following 24-h stimulation with TNF-α. ≠ p
    Vcam 1, supplied by rdi research diagnostics, used in various techniques. Bioz Stars score: 92/100, based on 17 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Pharmingen vcam 1
    Mechanisms of lymphocyte infiltration and localization of virus in pediatric lymphocytic interstitial pneumonia. A: Longitudinal section of a vein showing <t>VCAM-1</t> expression and associated mononuclear cell infiltrates. IP for VCAM-1 (clone 51–10C9, PharMingen); magnification, ×100. The inset shows a transverse section of a venule and demonstrates the uniform expression of endothelial VCAM-1. IP for VCAM-1; magnification, ×400. B: Perivenular aggregation of CD8 + T lymphocytes in areas of interstitial collapse. IP for CD8 (clone C8/144B, DAKO); magnification, ×400. C: Localization of cells harboring HIV mRNA ( arrows ) within a thin-walled vein. 125 I ISH for HIV-1 mRNA; magnification, ×100. D: Expression of CMV early gene RNA in cytomegaloid cells ( arrows ) adjacent to a vein and in a region of interstitial collapse and lymphocyte infiltration. ISH αDIG-AP for EBV EBER-1 RNA; magnification, ×400. E: Expression of KSHV latent gene transcripts in cells associated with follicular aggregation. ISH αDIG-AP for KSHV T0.7 RNA; magnification, ×400.
    Vcam 1, supplied by Pharmingen, used in various techniques. Bioz Stars score: 92/100, based on 161 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    SouthernBiotech vcam 1
    Outgrowth endothelial cell morphology and phenotype. Outgrowth cells have typical endothelial morphology ( a ). The remaining parts ( b – e ) show, as labeled at the bottom of each, constitutive and activated phenotype by immunofluorescent staining. Outgrowth endothelial cells incorporated acetylated LDL and were positive for vWF. They were negative for <t>VCAM-1</t> but expressed it upon stimulation.
    Vcam 1, supplied by SouthernBiotech, used in various techniques. Bioz Stars score: 92/100, based on 23 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Abcam vcam 1
    Results of gene expression by Semi-quantitative RT-PCR. Representative gel images of E-sel, ICAM-1, <t>VCAM-1</t> and MCP- 1 mRNA expression of the second portion of rabbit aortas. β-actin was used as an internal control and PCR products were visualized by ethidium bromide staining. The data in the bar graph are quantified ratios of the signal for E-sel, ICAM-1, VCAM-1 and MCP-1 to that for β-actin with the control samples set at 100%. Results are expressed as mean± SD (n = 6). ** p
    Vcam 1, supplied by Abcam, used in various techniques. Bioz Stars score: 94/100, based on 1152 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/vcam 1/product/Abcam
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    91
    Biorbyt vcam 1
    Bilirubin suppresses cellular  ROS  generation following activation of  VCAM ‐1 or  ICAM ‐1.  TNF ‐α‐activated  HUVEC  monolayers were incubated with anti‐ VCAM ‐1 (left panels, α VCAM ‐1; 10 μg/mL) or anti‐ ICAM ‐1 (right panels, α ICAM ‐1; 10 μg/mL) for 30 minutes and then loaded with dihydrorhodamine. Adhesion molecule activation was triggered by the addition of a cross‐linking antibody and  ROS  generation quantified by confocal microscopy. Upper panels display representative time‐lapse images of nonstimulated and antibody‐activated cells treated with 20 μmol/L of bilirubin ( BR ) or the bilirubin vehicle (Veh). Scale bars represent 100 μm. Lower panels plot the time‐dependent changes in fluorescence intensity following  VCAM ‐1 (left panel) or  ICAM ‐1 (right panel) activation (squares), in the presence (white symbols) or absence (black symbols) of bilirubin. Cells that were not treated with cross‐linking antibodies (nonstimulated; circles) serve as control, with curves reflecting mean fluorescence intensity (±SEM) expressed relative to maximal activation at 60 minutes (n=3 sets of experiments). * P
    Vcam 1, supplied by Biorbyt, used in various techniques. Bioz Stars score: 91/100, based on 79 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Cambridge Bioscience vcam 1
    MPIO-contrast volume reflects <t>VCAM-1</t> mRNA expression in tissue. VCAM-1 mRNA expression was 12-fold higher in clamped kidneys versus sham operated kidneys (n = 5, P
    Vcam 1, supplied by Cambridge Bioscience, used in various techniques. Bioz Stars score: 92/100, based on 77 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Effects of ImKTx88 on ICAM-1 and VCAM-1 expression. a Representative immunohistochemistry of ICAM-1 and VCAM-1 expression in the cerebellar white matter of EAE-induced and ImKTx88 treated rats. The scale bar represents 100 µm. Staining was quantified by mean of the integrated optical density (IOD) (5 random images per section and n = 3). b A representative western blot of cerebellum homogenates and quantification analysis revealed that the levels of ICAM-1 and VCAM-1 were significantly increased in EAE-induced rats, and ImKTx88 could decrease the levels of ICAM-1 and VCAM-1 (n = 5–12). GAPDH was used as an internal loading control. Data represent the mean ± SEM, * P

    Journal: Cell & Bioscience

    Article Title: Kv1.3 channel blocker (ImKTx88) maintains blood–brain barrier in experimental autoimmune encephalomyelitis

    doi: 10.1186/s13578-017-0158-2

    Figure Lengend Snippet: Effects of ImKTx88 on ICAM-1 and VCAM-1 expression. a Representative immunohistochemistry of ICAM-1 and VCAM-1 expression in the cerebellar white matter of EAE-induced and ImKTx88 treated rats. The scale bar represents 100 µm. Staining was quantified by mean of the integrated optical density (IOD) (5 random images per section and n = 3). b A representative western blot of cerebellum homogenates and quantification analysis revealed that the levels of ICAM-1 and VCAM-1 were significantly increased in EAE-induced rats, and ImKTx88 could decrease the levels of ICAM-1 and VCAM-1 (n = 5–12). GAPDH was used as an internal loading control. Data represent the mean ± SEM, * P

    Article Snippet: Blots were incubated overnight with the primary antibodies as follows: occludin (1:1000, Abcam, Cambridge, MA, USA); ZO-1 (1:500, Invitrogen, Carlsbad, CA, USA); claudin-5 (1:500, Abclonal, Woburn, MA, USA); ICAM-1(1:1000, Abclonal, Woburn, MA, USA); VCAM-1 (1:1000, Abclonal, Woburn, MA, USA); Ang-1 (1:500, Abclonal, Woburn, MA, USA); Tie-2 (1:500, Abclonal, Woburn, MA, USA); GAPDH (1:5000, Abclonal, Woburn, MA, USA).

    Techniques: Expressing, Immunohistochemistry, Staining, Western Blot

    Characterization of VCAM-1 + macrophages in the CHT. a, Transgenic Tg(kdrl:eGFP) embryos, stained with an anti-VCAM-1 antibody (magenta, arrows), show that the VCAM-1 + macrophage first appeared in the CHT at 32 h.p.f. b, Tg(kdrl:eGFP) embryos in the vcam1 cas011 , itga4 cas005 or runx1 w84x mutant background are stained with an anti-VCAM-1 antibody (magenta, white arrows) at 54 h.p.f. Signals in itga4 cas005 and runx1 w84x are similar to that in wild-type siblings, whereas there is almost no detectable signal in vcam1 cas011 mutants. c, Schematic diagrams (left) and confocal imaging (right) of VCAM-1 + macrophages (labelled with Alexa Fluor 647 dye-conjugated anti-VCAM-1 antibody by intravascular injection) that patrol the CHT of wild-type embryos. VCAM-1 + macrophages were mainly located intravascularly ( > 91%) with round or unpolarized cell morphology ( > 84%). Cross indicates the original position of VCAM-1 + . DA, dorsal aorta; VC, venous capillaries. Scale bars, 50 μm ( a, b ) and 20 μm ( c ).

    Journal: Nature

    Article Title: VCAM-1+ macrophages guide the homing of HSPCs to a vascular niche

    doi: 10.1038/s41586-018-0709-7

    Figure Lengend Snippet: Characterization of VCAM-1 + macrophages in the CHT. a, Transgenic Tg(kdrl:eGFP) embryos, stained with an anti-VCAM-1 antibody (magenta, arrows), show that the VCAM-1 + macrophage first appeared in the CHT at 32 h.p.f. b, Tg(kdrl:eGFP) embryos in the vcam1 cas011 , itga4 cas005 or runx1 w84x mutant background are stained with an anti-VCAM-1 antibody (magenta, white arrows) at 54 h.p.f. Signals in itga4 cas005 and runx1 w84x are similar to that in wild-type siblings, whereas there is almost no detectable signal in vcam1 cas011 mutants. c, Schematic diagrams (left) and confocal imaging (right) of VCAM-1 + macrophages (labelled with Alexa Fluor 647 dye-conjugated anti-VCAM-1 antibody by intravascular injection) that patrol the CHT of wild-type embryos. VCAM-1 + macrophages were mainly located intravascularly ( > 91%) with round or unpolarized cell morphology ( > 84%). Cross indicates the original position of VCAM-1 + . DA, dorsal aorta; VC, venous capillaries. Scale bars, 50 μm ( a, b ) and 20 μm ( c ).

    Article Snippet: For live labelling of usher cells, VCAM-1 antibody (Abclonal) was conjugated with Alexa Fluor 647 dye and purified by Microscale Protein Labelling Kit (Invitrogen, A30009).

    Techniques: Transgenic Assay, Staining, Mutagenesis, Imaging, Injection

    Anti-VCAM-1 647 antibody labels usher cells without disrupting definitive haematopoiesis. a, Injection of 1 nl (0.4 ng) of anti-VCAM-1 647 antibody labels usher cells (arrows) in wildtype and itga4 cas010 mutants in the Tg(kdrl:eGFP) background, whereas injection of either control (non-specific) IgG 647 antibody into wildtype cells or anti-VCAM-1 647 antibody into vcam1 cas011 mutants in the Tg(kdrl:eGFP) background did not label any cells in the retention hotspots. Asterisk indicates nonspecific labelling on a chromatophore in the CHT. b, Anti-VCAM-1 647 injection marginally influence definitive haematopoiesis. Statistical analysis shows the percentage of the three types of definitive haematopoietic phenotype in nine different conditions, including wild-type embryos without injection (#1), with 10 nl vehicle (#2) or 10 nl 0.4 mg ml −1 IgG 647 injection (#3), itga4 cas005 mutants (#4) or vcam1 cas011 mutants (#5) without injection, and wild-type embryos with 1–10 nl 0.4 mg ml −1 anti-VCAM-1 647 injection (#6-#9). c, d, Live imaging of HSPCs ( c ) or WISH analysis of the myb probe at 60 h.p.f. ( d ) of the wild-type CHT after vehicle or 1 nl anti-VCAM-1 647 antibody injection. e, Schematic diagrams (left) and confocal imaging (right) show VCAM-1 + cells patrolling on a small scale in the CHT in itga4 cas010 mutant embryos. Cross indicates the original position at the initial time point. f, Statistical analysis of the duration of the interaction between HSPCs and usher cells, the HSPC retention time, the diameter of vessels and the dorsal-ventral relative location for HSPC retention in pre-type I (type 0), pre-type II (type 0), type I and type II. Duration, pre-I vs pre-II: ***P = 0.0001, t = 4.25, df = 41; duration, pre-I vs I: ****P

    Journal: Nature

    Article Title: VCAM-1+ macrophages guide the homing of HSPCs to a vascular niche

    doi: 10.1038/s41586-018-0709-7

    Figure Lengend Snippet: Anti-VCAM-1 647 antibody labels usher cells without disrupting definitive haematopoiesis. a, Injection of 1 nl (0.4 ng) of anti-VCAM-1 647 antibody labels usher cells (arrows) in wildtype and itga4 cas010 mutants in the Tg(kdrl:eGFP) background, whereas injection of either control (non-specific) IgG 647 antibody into wildtype cells or anti-VCAM-1 647 antibody into vcam1 cas011 mutants in the Tg(kdrl:eGFP) background did not label any cells in the retention hotspots. Asterisk indicates nonspecific labelling on a chromatophore in the CHT. b, Anti-VCAM-1 647 injection marginally influence definitive haematopoiesis. Statistical analysis shows the percentage of the three types of definitive haematopoietic phenotype in nine different conditions, including wild-type embryos without injection (#1), with 10 nl vehicle (#2) or 10 nl 0.4 mg ml −1 IgG 647 injection (#3), itga4 cas005 mutants (#4) or vcam1 cas011 mutants (#5) without injection, and wild-type embryos with 1–10 nl 0.4 mg ml −1 anti-VCAM-1 647 injection (#6-#9). c, d, Live imaging of HSPCs ( c ) or WISH analysis of the myb probe at 60 h.p.f. ( d ) of the wild-type CHT after vehicle or 1 nl anti-VCAM-1 647 antibody injection. e, Schematic diagrams (left) and confocal imaging (right) show VCAM-1 + cells patrolling on a small scale in the CHT in itga4 cas010 mutant embryos. Cross indicates the original position at the initial time point. f, Statistical analysis of the duration of the interaction between HSPCs and usher cells, the HSPC retention time, the diameter of vessels and the dorsal-ventral relative location for HSPC retention in pre-type I (type 0), pre-type II (type 0), type I and type II. Duration, pre-I vs pre-II: ***P = 0.0001, t = 4.25, df = 41; duration, pre-I vs I: ****P

    Article Snippet: For live labelling of usher cells, VCAM-1 antibody (Abclonal) was conjugated with Alexa Fluor 647 dye and purified by Microscale Protein Labelling Kit (Invitrogen, A30009).

    Techniques: Injection, Imaging, Mutagenesis

    Characterization of VCAM-1 + cells in the CHT. a, Generation of the vcaml mutant using the CRISPR-Cas9 technique. The alignment of wild-type (underlined) and mutated sequences is listed. The PAM sequence of gRNA is ‘GGG’ (in blue). Deletions are indicated by dashes. b, According to the stop codon in the genome, SMART software was used to predict the structure of the wild-type vcaml and vcam1 cas011 presumed protein. The molecular sizes of the presumed protein are indicated. c, Live imaging of the CHT at 54 h.p.f. shows retention defects in vcam1 cas011 mutants. Representative images show that most HSPCs resided within the CHT (white arrows) in wild-type siblings (top), whereas these cells went through quickly in vcam1 cas011 ). d, WISH analysis of myb expression in the CHT of wild-type and vcam1 cas011 embryos at 72 h.p.f. e, f, The definitive haematopoiesis is defective in vcam1 cas011 mutant zebrafish embryos. e, The bright-field images of wild-type and vcam1 cas011 embryos show no obvious morphological difference at 72 h.p.f. f, WISH results of hbael.1, mpx and lyz expression in wild-type and vcam1 cas011 mutant embryos at 72 h.p.f. Arrows indicate the comparable position in wild-type (black) or vcam1 cas011 (red) embryos. g, Magnified views showed VCAM-1 was mainly expressed in individual cells (white arrow) but weakly expressed on the venous endothelial cells (yellow arrowheads). h, After photoconversion, Tg(kdrl:Dendra2) embryos are stained with anti-VCAM-1 (magenta, white arrow). The yellow arrowhead denotes an HSPC. i, Tg(cxcl12a:DsRed,kdrl:eGFP) transgenic embryos are stained with anti-VCAM-1 (magenta, white arrow) and anti-DsRed (red, yellow arrowhead). j, Tg(tcf:eGFP,kdrl:mCherry) transgenic embryos are stained with anti-VCAM-1 (magenta, white arrows) and anti-GFP (green, yellow arrowheads). Scale bars, 50 μm ( c ), 20 μm ( g ) and 10 μm ( h, i ).

    Journal: Nature

    Article Title: VCAM-1+ macrophages guide the homing of HSPCs to a vascular niche

    doi: 10.1038/s41586-018-0709-7

    Figure Lengend Snippet: Characterization of VCAM-1 + cells in the CHT. a, Generation of the vcaml mutant using the CRISPR-Cas9 technique. The alignment of wild-type (underlined) and mutated sequences is listed. The PAM sequence of gRNA is ‘GGG’ (in blue). Deletions are indicated by dashes. b, According to the stop codon in the genome, SMART software was used to predict the structure of the wild-type vcaml and vcam1 cas011 presumed protein. The molecular sizes of the presumed protein are indicated. c, Live imaging of the CHT at 54 h.p.f. shows retention defects in vcam1 cas011 mutants. Representative images show that most HSPCs resided within the CHT (white arrows) in wild-type siblings (top), whereas these cells went through quickly in vcam1 cas011 ). d, WISH analysis of myb expression in the CHT of wild-type and vcam1 cas011 embryos at 72 h.p.f. e, f, The definitive haematopoiesis is defective in vcam1 cas011 mutant zebrafish embryos. e, The bright-field images of wild-type and vcam1 cas011 embryos show no obvious morphological difference at 72 h.p.f. f, WISH results of hbael.1, mpx and lyz expression in wild-type and vcam1 cas011 mutant embryos at 72 h.p.f. Arrows indicate the comparable position in wild-type (black) or vcam1 cas011 (red) embryos. g, Magnified views showed VCAM-1 was mainly expressed in individual cells (white arrow) but weakly expressed on the venous endothelial cells (yellow arrowheads). h, After photoconversion, Tg(kdrl:Dendra2) embryos are stained with anti-VCAM-1 (magenta, white arrow). The yellow arrowhead denotes an HSPC. i, Tg(cxcl12a:DsRed,kdrl:eGFP) transgenic embryos are stained with anti-VCAM-1 (magenta, white arrow) and anti-DsRed (red, yellow arrowhead). j, Tg(tcf:eGFP,kdrl:mCherry) transgenic embryos are stained with anti-VCAM-1 (magenta, white arrows) and anti-GFP (green, yellow arrowheads). Scale bars, 50 μm ( c ), 20 μm ( g ) and 10 μm ( h, i ).

    Article Snippet: For live labelling of usher cells, VCAM-1 antibody (Abclonal) was conjugated with Alexa Fluor 647 dye and purified by Microscale Protein Labelling Kit (Invitrogen, A30009).

    Techniques: Mutagenesis, CRISPR, Sequencing, Software, Imaging, Expressing, Staining, Transgenic Assay

    Live imaging analysis on VCAM-1 + usher cell-guided HSPCs retention. a, Schematic illustration (top left) shows that labelling of HSPCs (with photoconverted Dendra2, red) was performed at 36 h.p.f. in Tg(kdrl:Dendra2) embryos, followed by an anti-VCAM-1 647 . Representative images (bottom) show the interaction between HSPCs (red; yellow arrowheads) and VCAM-1 + usher cells (magenta; white arrows). VCAM-1 + ). The top right images show one slice, and the others show z -stacks. b, ). Scale bars, 20 μm ( a ) and 10 μm ( b ).

    Journal: Nature

    Article Title: VCAM-1+ macrophages guide the homing of HSPCs to a vascular niche

    doi: 10.1038/s41586-018-0709-7

    Figure Lengend Snippet: Live imaging analysis on VCAM-1 + usher cell-guided HSPCs retention. a, Schematic illustration (top left) shows that labelling of HSPCs (with photoconverted Dendra2, red) was performed at 36 h.p.f. in Tg(kdrl:Dendra2) embryos, followed by an anti-VCAM-1 647 . Representative images (bottom) show the interaction between HSPCs (red; yellow arrowheads) and VCAM-1 + usher cells (magenta; white arrows). VCAM-1 + ). The top right images show one slice, and the others show z -stacks. b, ). Scale bars, 20 μm ( a ) and 10 μm ( b ).

    Article Snippet: For live labelling of usher cells, VCAM-1 antibody (Abclonal) was conjugated with Alexa Fluor 647 dye and purified by Microscale Protein Labelling Kit (Invitrogen, A30009).

    Techniques: Imaging

    Distinct role of macrophages and venous endothelium VCAM-1 in HSPCs retention. a, Representative FISH confocal imaging of mfap4 (top), csf1ra (middle), spi1a (bottom) immunofluorescence with anti-VCAM-1 and anti-GFP antibodies indicates that VCAM-1 + ). b, The construction of the plasmid applied in c–f. c, Validation of the macrophage-specific cell-depletion system. Left, the number of HSPCs under the endothelial-to-haematopoietic transition (EHT) process in the AGM of Tg(mpeg1:Gal4,kdrl:Dendra2) transgenic embryos at 54 h.p.f. with MTZ treatment and with (#2) or without (#1) the Tg(UAS:NfsB-mCherry) background. P = 0.80, t = 0.25, df = 9. Right, live imaging of vessels (green) and macrophages (red) with or without MTZ treatment in the CHT of Tg(kdrl:Dendra2,mpeg1:Gal4,UAS :N fsB-mCherry) transgenic embryos showed that MTZ treatment could delete almost all mCherry + macrophages. d, Quantification of VCAM-1 + cells in the CHT, detected by anti-VCAM-1 immunofluorescence, in Tg(mpeg1:Gal4,kdrl:Dendra2) embryos at 54 h.p.f. with MTZ treatment and with (#2) or without (#1) a Tg(UAS:NfsB-mCherry) background, and in vcam1 cas011 mutants with Tol2-mediated transient transgenesis of vector (UAS:polyA) (#3) or UAS:vcam1 (#4) in a Tg(mpeg1:Gal4,kdrl:Dendra2) background. #1 vs #2: ****P

    Journal: Nature

    Article Title: VCAM-1+ macrophages guide the homing of HSPCs to a vascular niche

    doi: 10.1038/s41586-018-0709-7

    Figure Lengend Snippet: Distinct role of macrophages and venous endothelium VCAM-1 in HSPCs retention. a, Representative FISH confocal imaging of mfap4 (top), csf1ra (middle), spi1a (bottom) immunofluorescence with anti-VCAM-1 and anti-GFP antibodies indicates that VCAM-1 + ). b, The construction of the plasmid applied in c–f. c, Validation of the macrophage-specific cell-depletion system. Left, the number of HSPCs under the endothelial-to-haematopoietic transition (EHT) process in the AGM of Tg(mpeg1:Gal4,kdrl:Dendra2) transgenic embryos at 54 h.p.f. with MTZ treatment and with (#2) or without (#1) the Tg(UAS:NfsB-mCherry) background. P = 0.80, t = 0.25, df = 9. Right, live imaging of vessels (green) and macrophages (red) with or without MTZ treatment in the CHT of Tg(kdrl:Dendra2,mpeg1:Gal4,UAS :N fsB-mCherry) transgenic embryos showed that MTZ treatment could delete almost all mCherry + macrophages. d, Quantification of VCAM-1 + cells in the CHT, detected by anti-VCAM-1 immunofluorescence, in Tg(mpeg1:Gal4,kdrl:Dendra2) embryos at 54 h.p.f. with MTZ treatment and with (#2) or without (#1) a Tg(UAS:NfsB-mCherry) background, and in vcam1 cas011 mutants with Tol2-mediated transient transgenesis of vector (UAS:polyA) (#3) or UAS:vcam1 (#4) in a Tg(mpeg1:Gal4,kdrl:Dendra2) background. #1 vs #2: ****P

    Article Snippet: For live labelling of usher cells, VCAM-1 antibody (Abclonal) was conjugated with Alexa Fluor 647 dye and purified by Microscale Protein Labelling Kit (Invitrogen, A30009).

    Techniques: Fluorescence In Situ Hybridization, Imaging, Immunofluorescence, Plasmid Preparation, Transgenic Assay

    Distinct role of macrophages and venous endothelium VCAM-1 in HSPCs retention. a, Percentage of total HSPCs in four classified retention time zones in grouped wild-type siblings and vcam1 cas011 mutants (n = 3) at 50–60 h.p.f. b, Tg ( kdrl:eGFP ) embryos, stained with an anti-VCAM-1 antibody (magenta, arrows), show dorsal venous plexus distribution of individual VCAM-1 + cells in the CHT. c, Percentage of VCAM-1 + cells in the CHT scored by the distance to the nearest HSPC (edge to edge, n = 100). Most (86%; 86 out of 100 cells) VCAM-1 + cells were located within 7 μm of HSPCs (the average diameter of HSPCs is about 6.9 μm). Tg(kdrl:Dendra2) embryos with AGM photoconversion were stained with an anti-VCAM-1 antibody (magenta, white arrow). Yellow arrowheads denote HSPCs. d, The staining of Tg(mpeg1:eGFP) embryos with an anti-VCAM-1 antibody (magenta) shows that VCAM-1 + cells merge with mpeg1 + cells (green) in the CHT. White arrowheads denote VCAM-1 + GFP + double-positive cells; yellow arrowheads denote GFP single-positive cells. e, Live-imaging frame shots of HSPCs in macrophage-specific cell-depletion embryos from f . Time is in minutes:seconds. f, Percentage of total HSPCs in four classified retention time zones in grouped ( n = 3) Tg(mpeg1:Gal4,kdrl:Dendra2) embryos with MTZ treatment, with or without the Tg(UAS:NfsB-mCherry) background, at 50–60 h.p.f. g, Percentage of total HSPCs in four classified retention time zones in grouped (n = 3) vcam1 cas011 mutants in a Tg(mpeg1:Gal4,kdrl:Dendra2) background with transient transgenesis of either vector (UAS :polyA) or UAS:vcam1 . Scale bars, 50 μm ( b, d ), 20 μm ( e ) and 10 μm ( c ).

    Journal: Nature

    Article Title: VCAM-1+ macrophages guide the homing of HSPCs to a vascular niche

    doi: 10.1038/s41586-018-0709-7

    Figure Lengend Snippet: Distinct role of macrophages and venous endothelium VCAM-1 in HSPCs retention. a, Percentage of total HSPCs in four classified retention time zones in grouped wild-type siblings and vcam1 cas011 mutants (n = 3) at 50–60 h.p.f. b, Tg ( kdrl:eGFP ) embryos, stained with an anti-VCAM-1 antibody (magenta, arrows), show dorsal venous plexus distribution of individual VCAM-1 + cells in the CHT. c, Percentage of VCAM-1 + cells in the CHT scored by the distance to the nearest HSPC (edge to edge, n = 100). Most (86%; 86 out of 100 cells) VCAM-1 + cells were located within 7 μm of HSPCs (the average diameter of HSPCs is about 6.9 μm). Tg(kdrl:Dendra2) embryos with AGM photoconversion were stained with an anti-VCAM-1 antibody (magenta, white arrow). Yellow arrowheads denote HSPCs. d, The staining of Tg(mpeg1:eGFP) embryos with an anti-VCAM-1 antibody (magenta) shows that VCAM-1 + cells merge with mpeg1 + cells (green) in the CHT. White arrowheads denote VCAM-1 + GFP + double-positive cells; yellow arrowheads denote GFP single-positive cells. e, Live-imaging frame shots of HSPCs in macrophage-specific cell-depletion embryos from f . Time is in minutes:seconds. f, Percentage of total HSPCs in four classified retention time zones in grouped ( n = 3) Tg(mpeg1:Gal4,kdrl:Dendra2) embryos with MTZ treatment, with or without the Tg(UAS:NfsB-mCherry) background, at 50–60 h.p.f. g, Percentage of total HSPCs in four classified retention time zones in grouped (n = 3) vcam1 cas011 mutants in a Tg(mpeg1:Gal4,kdrl:Dendra2) background with transient transgenesis of either vector (UAS :polyA) or UAS:vcam1 . Scale bars, 50 μm ( b, d ), 20 μm ( e ) and 10 μm ( c ).

    Article Snippet: For live labelling of usher cells, VCAM-1 antibody (Abclonal) was conjugated with Alexa Fluor 647 dye and purified by Microscale Protein Labelling Kit (Invitrogen, A30009).

    Techniques: Staining, Imaging, Plasmid Preparation

    TNF-induced VCAM1 expression was decreased in TXNIP-deficient mouse aorta. After aortae from HcB-19 and control C3H mice were treated with TNF (15 ng/ml, 6 hours), vessel protein was harvested. Expression of VCAM1 and TXNIP was determined by immunoblotting

    Journal:

    Article Title: Fluid shear stress inhibits vascular inflammation by decreasing thioredoxin-interacting protein in endothelial cells

    doi: 10.1172/JCI200523001

    Figure Lengend Snippet: TNF-induced VCAM1 expression was decreased in TXNIP-deficient mouse aorta. After aortae from HcB-19 and control C3H mice were treated with TNF (15 ng/ml, 6 hours), vessel protein was harvested. Expression of VCAM1 and TXNIP was determined by immunoblotting

    Article Snippet: Antibody sources were Santa Cruz Biotechnology Inc. for VCAM1, ERK1/2, p38, eNOS, actin, and IκB-α; Cell Signaling Technology Inc. for phospho-ERK1/2, phospho-p38, JNK2, and ASK1; Promega Corp. for phospho-JNK; and American Diagnostica Inc. for human TRX1.

    Techniques: Expressing, Mouse Assay

    TXNIP siRNA inhibited TNF-induced VCAM1 expression in HUVECs. After HUVECs were transfected with either control or TXNIP siRNA, TNF-α (10 ng/ml) was added for 6 hours. VCAM1 expression was determined by immunoblotting from 3 independent experiments.

    Journal:

    Article Title: Fluid shear stress inhibits vascular inflammation by decreasing thioredoxin-interacting protein in endothelial cells

    doi: 10.1172/JCI200523001

    Figure Lengend Snippet: TXNIP siRNA inhibited TNF-induced VCAM1 expression in HUVECs. After HUVECs were transfected with either control or TXNIP siRNA, TNF-α (10 ng/ml) was added for 6 hours. VCAM1 expression was determined by immunoblotting from 3 independent experiments.

    Article Snippet: Antibody sources were Santa Cruz Biotechnology Inc. for VCAM1, ERK1/2, p38, eNOS, actin, and IκB-α; Cell Signaling Technology Inc. for phospho-ERK1/2, phospho-p38, JNK2, and ASK1; Promega Corp. for phospho-JNK; and American Diagnostica Inc. for human TRX1.

    Techniques: Expressing, Transfection

    Flow regulates TXNIP in ECs. Chronic exposure to normal flow decreases TXNIP expression, and this results in increased TRX binding to ASK1. This inhibits cytokine activation of the JNK-p38 pathway and prevents proinflammatory events such as VCAM1 expression.

    Journal:

    Article Title: Fluid shear stress inhibits vascular inflammation by decreasing thioredoxin-interacting protein in endothelial cells

    doi: 10.1172/JCI200523001

    Figure Lengend Snippet: Flow regulates TXNIP in ECs. Chronic exposure to normal flow decreases TXNIP expression, and this results in increased TRX binding to ASK1. This inhibits cytokine activation of the JNK-p38 pathway and prevents proinflammatory events such as VCAM1 expression.

    Article Snippet: Antibody sources were Santa Cruz Biotechnology Inc. for VCAM1, ERK1/2, p38, eNOS, actin, and IκB-α; Cell Signaling Technology Inc. for phospho-ERK1/2, phospho-p38, JNK2, and ASK1; Promega Corp. for phospho-JNK; and American Diagnostica Inc. for human TRX1.

    Techniques: Flow Cytometry, Expressing, Binding Assay, Activation Assay

    Direct effects of 12/15-LOX-derived metabolites on retinal vasculature. A: Fluorescein angiography (FA) of normal WT mice injected intravitreally with vehicle, as a control, or 12-HETE. One week later, FA was performed to evaluate changes of retinal vasculature. The relative fluorescence intensity of FA per mouse retina was calculated by the ImageJ software then normalized as a percentage to that of vehicle-injected control, which was arbitrarily set at 100%. B: Western blot of total retinal albumin among the studied groups followed by densitometric analysis. Ratio of the albumin band intensity relative to actin for the 12-HETE-injected group was compared with the vehicle-injected control, which was arbitrarily set at 1.0. C: Western blot analysis of retinal ICAM-1, VCAM-1, CD45, and NOX2 after intraocular injection with either 12-HETE (0.1 µM), or vehicle followed by densitometric analysis. Ratios of band intensities of ICAM-1, VCAM-1, CD45, and NOX2, respectively, relative to the actin for 12-HETE-injected group were compared with the vehicle-injected control, which was arbitrarily set at 1.0. Data shown for the comparison are the mean ± SD and representative of four to six mice studied in each group.

    Journal: Journal of Lipid Research

    Article Title: A lipidomic screen of hyperglycemia-treated HRECs links 12/15-Lipoxygenase to microvascular dysfunction during diabetic retinopathy via NADPH oxidase

    doi: 10.1194/jlr.M056069

    Figure Lengend Snippet: Direct effects of 12/15-LOX-derived metabolites on retinal vasculature. A: Fluorescein angiography (FA) of normal WT mice injected intravitreally with vehicle, as a control, or 12-HETE. One week later, FA was performed to evaluate changes of retinal vasculature. The relative fluorescence intensity of FA per mouse retina was calculated by the ImageJ software then normalized as a percentage to that of vehicle-injected control, which was arbitrarily set at 100%. B: Western blot of total retinal albumin among the studied groups followed by densitometric analysis. Ratio of the albumin band intensity relative to actin for the 12-HETE-injected group was compared with the vehicle-injected control, which was arbitrarily set at 1.0. C: Western blot analysis of retinal ICAM-1, VCAM-1, CD45, and NOX2 after intraocular injection with either 12-HETE (0.1 µM), or vehicle followed by densitometric analysis. Ratios of band intensities of ICAM-1, VCAM-1, CD45, and NOX2, respectively, relative to the actin for 12-HETE-injected group were compared with the vehicle-injected control, which was arbitrarily set at 1.0. Data shown for the comparison are the mean ± SD and representative of four to six mice studied in each group.

    Article Snippet: Soluble ICAM-1 and VCAM-1 as markers of endothelial activation .

    Techniques: Derivative Assay, Mouse Assay, Injection, Fluorescence, Software, Western Blot

    Effect of EPC Exosomal miR-126-3p and 5p on LPS-Induced HMVEC Target Expression and CLP-Induced Mortality Protein levels of VCAM1 (A) and HMGB1 (B) in HMVECs were measured by western blot. α-Tubulin served as an internal control. * p

    Journal: Molecular Therapy

    Article Title: Exosomes from Endothelial Progenitor Cells Improve the Outcome of a Murine Model of Sepsis

    doi: 10.1016/j.ymthe.2018.02.020

    Figure Lengend Snippet: Effect of EPC Exosomal miR-126-3p and 5p on LPS-Induced HMVEC Target Expression and CLP-Induced Mortality Protein levels of VCAM1 (A) and HMGB1 (B) in HMVECs were measured by western blot. α-Tubulin served as an internal control. * p

    Article Snippet: Primary antibodies including anti-HMGB1 (1:1,000; Cell Signaling, Boston, MA, USA) and anti-VCAM1 (1:500; Cell Signaling, Boston, MA, USA) were used.

    Techniques: Expressing, Western Blot

    DZ2002 reduced ICAM-1 and VCAM-1 in vivo and in vitro. a Western blot of ICAM-1 and VCAM-1 in BLM-induced SSc mice skin (DZ2002 = 50 mg/kg). b Survival rate of HMEC-1 at different concentrations of DZ2002 treatment. c Evaluation of mRNA levels of molecules associated with the upregulation of endothelial adhesion molecules in HMEC-1 by quantitative real-time reverse transcription-PCR, such as ICAM-1, VCAM-1, VEGF, bFGF, and ET-1. d Western blot of ICAM-1 and VCAM-1 in the HMEC-1 cells (TNF-α = 40 ng/ml, DZ2002 = 200 μM). e Representative photomicrographs showing the stained THP-1 cells on HMEC-1 cell layer (× 40, scale bar = 100 μm). The HMEC-1 cells were stimulated with TNF-a (40 ng/ml) with or without DZ2002 (200 μM). After 6 h incubation, those stained THP-1 cells adhered on HMEC-1 cell layer were counted. Mean ± SEM. * P

    Journal: Arthritis Research & Therapy

    Article Title: DZ2002 ameliorates fibrosis, inflammation, and vasculopathy in experimental systemic sclerosis models

    doi: 10.1186/s13075-019-2074-9

    Figure Lengend Snippet: DZ2002 reduced ICAM-1 and VCAM-1 in vivo and in vitro. a Western blot of ICAM-1 and VCAM-1 in BLM-induced SSc mice skin (DZ2002 = 50 mg/kg). b Survival rate of HMEC-1 at different concentrations of DZ2002 treatment. c Evaluation of mRNA levels of molecules associated with the upregulation of endothelial adhesion molecules in HMEC-1 by quantitative real-time reverse transcription-PCR, such as ICAM-1, VCAM-1, VEGF, bFGF, and ET-1. d Western blot of ICAM-1 and VCAM-1 in the HMEC-1 cells (TNF-α = 40 ng/ml, DZ2002 = 200 μM). e Representative photomicrographs showing the stained THP-1 cells on HMEC-1 cell layer (× 40, scale bar = 100 μm). The HMEC-1 cells were stimulated with TNF-a (40 ng/ml) with or without DZ2002 (200 μM). After 6 h incubation, those stained THP-1 cells adhered on HMEC-1 cell layer were counted. Mean ± SEM. * P

    Article Snippet: After blocking, the membranes were incubated with anti-TGF-β, anti-Smad3, anti-phospho-Smad3, anti-Smad4, anti-Smad7, anti-STAT1, anti-phospho-STAT1, anti-inducible NO synthase (iNOS), anti-Arginase 1 (Arg-1), anti-ICAM-1, and anti-VCAM-1 (all from Cell Signaling Technology, Beverly, MA, USA) (Mouse specific ICAM-1, Abcam, Cambridge, UK).

    Techniques: In Vivo, In Vitro, Western Blot, Mouse Assay, Polymerase Chain Reaction, Staining, Incubation

    Effect of the Taraxacum officinale (TO) methanol extract on LPS-induced VCAM-1 and MCP-1 expression. HUVECs were pretreated with TO extract (100 μg/ml) for 1 h and then incubated with 1 μg/ml LPS for 24 h. a Total cell lysate was prepared, the proteins were separated by SDS-PAGE, and the VCAM-1 levels were assessed by western blot analysis. b The mRNA levels were normalized to that of a housekeeping gene (18S rRNA) as well as the vehicle-only-treated control, and the 2 -ΔΔCt for each mRNA is reported. Results shown are the mean ± SE of 3 independent experiments. ** p

    Journal: BMC Complementary and Alternative Medicine

    Article Title: Anti-inflammatory evaluation of the methanolic extract of Taraxacum officinale in LPS-stimulated human umbilical vein endothelial cells

    doi: 10.1186/s12906-017-2022-7

    Figure Lengend Snippet: Effect of the Taraxacum officinale (TO) methanol extract on LPS-induced VCAM-1 and MCP-1 expression. HUVECs were pretreated with TO extract (100 μg/ml) for 1 h and then incubated with 1 μg/ml LPS for 24 h. a Total cell lysate was prepared, the proteins were separated by SDS-PAGE, and the VCAM-1 levels were assessed by western blot analysis. b The mRNA levels were normalized to that of a housekeeping gene (18S rRNA) as well as the vehicle-only-treated control, and the 2 -ΔΔCt for each mRNA is reported. Results shown are the mean ± SE of 3 independent experiments. ** p

    Article Snippet: Antibodies against VCAM-1, intercellular adhesion molecule-1 (ICAM-1), β-actin, NF-κB p65, inhibitor of NF-κB alpha (IκBα), and phospho-IκBα were obtained from Cell Signaling Technology (Danvers, MA, USA).

    Techniques: Expressing, Incubation, SDS Page, Western Blot

    Immunofluorescence analysis of BBB endothelial expression of VCAM-1 and E-selectin (A) and PECAM1 (B), following exposure to CSEs from 1R5F, 3R4F, NF and ultralow nicotine cigarettes. Immunofluorescence analysis of PECAM1 was confirmed by WB of corresponding membrane fractions. (C) Release of proinflammatory cytokines IL-6 was up-regulated in endothelial cultures exposed to NF and ultralow nicotine CSE while MMP-2 levels were increased by CSE from 3R4F and NF but not ultralow nicotine. n = 3 biological replicates, *p

    Journal: BMC Neuroscience

    Article Title: Oxidative and pro-inflammatory impact of regular and denicotinized cigarettes on blood brain barrier endothelial cells: is smoking reduced or nicotine-free products really safe?

    doi: 10.1186/1471-2202-15-51

    Figure Lengend Snippet: Immunofluorescence analysis of BBB endothelial expression of VCAM-1 and E-selectin (A) and PECAM1 (B), following exposure to CSEs from 1R5F, 3R4F, NF and ultralow nicotine cigarettes. Immunofluorescence analysis of PECAM1 was confirmed by WB of corresponding membrane fractions. (C) Release of proinflammatory cytokines IL-6 was up-regulated in endothelial cultures exposed to NF and ultralow nicotine CSE while MMP-2 levels were increased by CSE from 3R4F and NF but not ultralow nicotine. n = 3 biological replicates, *p

    Article Snippet: Materials and reagents The antibodies used in this study were obtained from the following sources: Rabbit anti-ZO-1 (#8193), rabbit anti-claudin-3 (#341700), rabbit anti-VE-cadherin (#D87F2), rabbit anti-VCAM-1 (#12367), mouse PECAM-1 (#89C2) from Cell Signaling Technology (Danvers, MA, USA); mouse anti-E-selectin (#S 9555), β-actin (#A5441) from Sigma-Aldrich (St. Louis, MO, USA); donkey anti-rabbit (#NA934) and sheep anti-mouse (#NA931) HRP-linked secondary antibodies from GE Healthcare (Piscataway, NJ, USA); mouse anti-claudin 5 (#35-2500), goat anti-rabbit (#A11008) and anti-mouse (#A21422) conjugated to Alexa Fluor® 488 and 555 from Invitrogen (Camarillo, CA, USA).

    Techniques: Immunofluorescence, Expressing, Western Blot

    Gene ontology and localization of EC subtypes elucidates known functions and confirms divergence of clusters 2 and 4. ( A ) Circos plot shows gene expression and gene ontology (GO) overlap between distinct clusters of pulmonary ECs. A purple line connecting two clusters indicates expression of the same gene in both clusters, while a blue line connecting two clusters indicates expression of different genes found within the same GO category in each cluster. ( B ) GO biological process enrichment performed for each cluster and displayed in heatmap format demonstrates expression of genes associated with different biological processes in each cluster, including some overlap between clusters. Cluster 2 (green box) shares enrichment of some processes related to angiogenesis and blood vessel development with cluster 0 (blue box) but is distinct in its enrichment of genes related to vasculogenesis. ( C ) In-depth analysis of gene expression in cluster four indicates that this cluster likely represents macrovascular ECs (maECs) with high expression of Vwf and Vcam1 . IHC indicates that these proteins localize mainly to the large vessel endothelium. White arrowhead demonstrates Vcam1 located in nearby mesenchymal cells. ( D ) Clusters 0, 1, and three represent a heterogeneous population of microvascular ECs (miECs) with high expression of Gpihbp1 and Plvap . IHC indicates that these proteins localize to the alveolar capillary plexus endothelium. Yellow arrowhead demonstrates Plvap present in the large vessel endothelium. ( E ) Cluster two represents an as-yet uncharacterized population of ECs that localize to the alveolar region and express surface marker Cd34 at a higher level from that in other ECs. These cells also express high levels of Car4 . CD34 and Car4 proteins localize to the alveolar space, indicating a similar spatial distribution of cells in cluster two to that of miECs. v, vessel; a, alveolar space; scale bars in ( C )-( E ), 20 microns.

    Journal: eLife

    Article Title: Defining the role of pulmonary endothelial cell heterogeneity in the response to acute lung injury

    doi: 10.7554/eLife.53072

    Figure Lengend Snippet: Gene ontology and localization of EC subtypes elucidates known functions and confirms divergence of clusters 2 and 4. ( A ) Circos plot shows gene expression and gene ontology (GO) overlap between distinct clusters of pulmonary ECs. A purple line connecting two clusters indicates expression of the same gene in both clusters, while a blue line connecting two clusters indicates expression of different genes found within the same GO category in each cluster. ( B ) GO biological process enrichment performed for each cluster and displayed in heatmap format demonstrates expression of genes associated with different biological processes in each cluster, including some overlap between clusters. Cluster 2 (green box) shares enrichment of some processes related to angiogenesis and blood vessel development with cluster 0 (blue box) but is distinct in its enrichment of genes related to vasculogenesis. ( C ) In-depth analysis of gene expression in cluster four indicates that this cluster likely represents macrovascular ECs (maECs) with high expression of Vwf and Vcam1 . IHC indicates that these proteins localize mainly to the large vessel endothelium. White arrowhead demonstrates Vcam1 located in nearby mesenchymal cells. ( D ) Clusters 0, 1, and three represent a heterogeneous population of microvascular ECs (miECs) with high expression of Gpihbp1 and Plvap . IHC indicates that these proteins localize to the alveolar capillary plexus endothelium. Yellow arrowhead demonstrates Plvap present in the large vessel endothelium. ( E ) Cluster two represents an as-yet uncharacterized population of ECs that localize to the alveolar region and express surface marker Cd34 at a higher level from that in other ECs. These cells also express high levels of Car4 . CD34 and Car4 proteins localize to the alveolar space, indicating a similar spatial distribution of cells in cluster two to that of miECs. v, vessel; a, alveolar space; scale bars in ( C )-( E ), 20 microns.

    Article Snippet: Immunohistochemistry was used to recognize the antigens of various ECs, using the following antibodies: CD31 (Rat, HistoBioTec DIA-310), CD31 (Rabbit, Thermo Fisher MA5-16337), Vwf (Rabbit, Dako A0082), Vcam1 (Rat, eBioscience 14–1061), Gpihbp1 (Rabbit, Thermo Fisher PA1-16976), Plvap (Rat, BioRad MCA2539T), CD34 (Rat, PharMingen 553731), Car4 (Rat, R and D Systems MAB2414), Ki67 (Rabbit, Abcam ab16667), phospho-histone H3 (pHH3, mouse, Cell Signaling Technologies 9706), Sftpc (Goat, Santa Cruz Biotechnology sc-7706), and Hopx (Mouse, Santa Cruz Biotechnology sc-398703).

    Techniques: Expressing, Immunohistochemistry, Marker

    Proliferating ECs express general EC marker genes, but do not express genes that are highly expressed in Car4 -high ECs. ( A ) UMAP dimension reduction of H1N1 injury scRNA-seq dataset, with red box enclosing EC clusters and representing areas of increased digital zoom displayed in ( B )-( I ). Black circle demarcates proliferating EC cluster. Proliferating ECs express genes highly expressed in all ECs or in miECs, such as ( B ) Cd31 , ( C ) Gpihbp1 , and ( D ) Plvap . However, they express moderate levels of ( E ) Cd34 and do not express other genes that are highly expressed in Car4 -high ECs, such as ( F ) Kdr or ( G ) Ednrb . Proliferating ECs do not express genes enriched in maECs, such as ( H ) Vcam1 or ( I ) Vwf.

    Journal: eLife

    Article Title: Defining the role of pulmonary endothelial cell heterogeneity in the response to acute lung injury

    doi: 10.7554/eLife.53072

    Figure Lengend Snippet: Proliferating ECs express general EC marker genes, but do not express genes that are highly expressed in Car4 -high ECs. ( A ) UMAP dimension reduction of H1N1 injury scRNA-seq dataset, with red box enclosing EC clusters and representing areas of increased digital zoom displayed in ( B )-( I ). Black circle demarcates proliferating EC cluster. Proliferating ECs express genes highly expressed in all ECs or in miECs, such as ( B ) Cd31 , ( C ) Gpihbp1 , and ( D ) Plvap . However, they express moderate levels of ( E ) Cd34 and do not express other genes that are highly expressed in Car4 -high ECs, such as ( F ) Kdr or ( G ) Ednrb . Proliferating ECs do not express genes enriched in maECs, such as ( H ) Vcam1 or ( I ) Vwf.

    Article Snippet: Immunohistochemistry was used to recognize the antigens of various ECs, using the following antibodies: CD31 (Rat, HistoBioTec DIA-310), CD31 (Rabbit, Thermo Fisher MA5-16337), Vwf (Rabbit, Dako A0082), Vcam1 (Rat, eBioscience 14–1061), Gpihbp1 (Rabbit, Thermo Fisher PA1-16976), Plvap (Rat, BioRad MCA2539T), CD34 (Rat, PharMingen 553731), Car4 (Rat, R and D Systems MAB2414), Ki67 (Rabbit, Abcam ab16667), phospho-histone H3 (pHH3, mouse, Cell Signaling Technologies 9706), Sftpc (Goat, Santa Cruz Biotechnology sc-7706), and Hopx (Mouse, Santa Cruz Biotechnology sc-398703).

    Techniques: Marker

    Total and CD11b lo macrophage subsets are greatly reduced before and during E-stress response in Spi-C–deficient and VCAM-1–deficient mice. Numbers of CD11b lo (lighter shades) and CD11b hi (darker shades) macrophages, as well as VCAM-1 expression

    Journal: Blood

    Article Title: The macrophage contribution to stress erythropoiesis: when less is enough

    doi: 10.1182/blood-2016-05-714527

    Figure Lengend Snippet: Total and CD11b lo macrophage subsets are greatly reduced before and during E-stress response in Spi-C–deficient and VCAM-1–deficient mice. Numbers of CD11b lo (lighter shades) and CD11b hi (darker shades) macrophages, as well as VCAM-1 expression

    Article Snippet: For macrophage evaluation, we used VCAM-1–Alexa 488 (clone 429; BioLegend, San Diego, CA), CD11b-phycoerythrin (PE) (BioLegend), and F4/80-allophycocyanin (APC) (eBioscience, San Diego, CA); in some experiments, CD169-PE (clone 3D6.112; BioLegend) was used in combination with VCAM-1, F4/80, and CD11b mAbs; to assess cycling macrophages, Ki-67–fluorescein isothiocyanate (FITC) (BD Biosciences, San Jose, CA) was used together with F4/80, CD11b, and VCAM-1 mAbs according to the manufacturer’s instructions.

    Techniques: Mouse Assay, Expressing

    Macrophage subsets in WT mice at steady state and during response to stress. (A) Morphology and expression of F4/80, CD11b, and VCAM-1 was analyzed in EI preparations isolated from spleens of PHZ-treated or Epo-treated mice on day 4; Hema3 stain: note

    Journal: Blood

    Article Title: The macrophage contribution to stress erythropoiesis: when less is enough

    doi: 10.1182/blood-2016-05-714527

    Figure Lengend Snippet: Macrophage subsets in WT mice at steady state and during response to stress. (A) Morphology and expression of F4/80, CD11b, and VCAM-1 was analyzed in EI preparations isolated from spleens of PHZ-treated or Epo-treated mice on day 4; Hema3 stain: note

    Article Snippet: For macrophage evaluation, we used VCAM-1–Alexa 488 (clone 429; BioLegend, San Diego, CA), CD11b-phycoerythrin (PE) (BioLegend), and F4/80-allophycocyanin (APC) (eBioscience, San Diego, CA); in some experiments, CD169-PE (clone 3D6.112; BioLegend) was used in combination with VCAM-1, F4/80, and CD11b mAbs; to assess cycling macrophages, Ki-67–fluorescein isothiocyanate (FITC) (BD Biosciences, San Jose, CA) was used together with F4/80, CD11b, and VCAM-1 mAbs according to the manufacturer’s instructions.

    Techniques: Mouse Assay, Expressing, Isolation, Staining

    Erythroid responses to PHZ challenge in Spi-C–deficient and VCAM-1–deficient mice. (A) Cellularity and total number of Ebs in the femur (left panels) and spleen (right panels) of respective controls for each group of mice (red bars), Spi-C–deficient

    Journal: Blood

    Article Title: The macrophage contribution to stress erythropoiesis: when less is enough

    doi: 10.1182/blood-2016-05-714527

    Figure Lengend Snippet: Erythroid responses to PHZ challenge in Spi-C–deficient and VCAM-1–deficient mice. (A) Cellularity and total number of Ebs in the femur (left panels) and spleen (right panels) of respective controls for each group of mice (red bars), Spi-C–deficient

    Article Snippet: For macrophage evaluation, we used VCAM-1–Alexa 488 (clone 429; BioLegend, San Diego, CA), CD11b-phycoerythrin (PE) (BioLegend), and F4/80-allophycocyanin (APC) (eBioscience, San Diego, CA); in some experiments, CD169-PE (clone 3D6.112; BioLegend) was used in combination with VCAM-1, F4/80, and CD11b mAbs; to assess cycling macrophages, Ki-67–fluorescein isothiocyanate (FITC) (BD Biosciences, San Jose, CA) was used together with F4/80, CD11b, and VCAM-1 mAbs according to the manufacturer’s instructions.

    Techniques: Mouse Assay

    Dynamics of macrophage subsets CD11b lo (lighter shades) and CD11b hi (darker shades) in response to aCD24 antibody treatment. WT (red bars) and VCAM-1 −/− blue bars) mice were treated with aCD24 antibody or control immunoglobulin G (see

    Journal: Blood

    Article Title: The macrophage contribution to stress erythropoiesis: when less is enough

    doi: 10.1182/blood-2016-05-714527

    Figure Lengend Snippet: Dynamics of macrophage subsets CD11b lo (lighter shades) and CD11b hi (darker shades) in response to aCD24 antibody treatment. WT (red bars) and VCAM-1 −/− blue bars) mice were treated with aCD24 antibody or control immunoglobulin G (see

    Article Snippet: For macrophage evaluation, we used VCAM-1–Alexa 488 (clone 429; BioLegend, San Diego, CA), CD11b-phycoerythrin (PE) (BioLegend), and F4/80-allophycocyanin (APC) (eBioscience, San Diego, CA); in some experiments, CD169-PE (clone 3D6.112; BioLegend) was used in combination with VCAM-1, F4/80, and CD11b mAbs; to assess cycling macrophages, Ki-67–fluorescein isothiocyanate (FITC) (BD Biosciences, San Jose, CA) was used together with F4/80, CD11b, and VCAM-1 mAbs according to the manufacturer’s instructions.

    Techniques: Mouse Assay

    Immunohistochemical staining for F4/80-positive monocytes-derived macrophages and adhesion molecule VCAM-1 in aortic cross-sections Representative photomicrographs of immunohistochemical staining for F4/80-positive monocytes-derived macrophages (Fig. 6A) and VCAM-1 (Fig. 6B). Quantitative analysis of F4/80 (Fig. 6C) and VCAM-1 (Fig. 6D). Arrows indicate typical positive stained regions and original magnification is 40×. T, TNF-α; T +G, TNF-α + genistein; Data are expressed as mean ± SEM, n=5, *, p

    Journal: International journal of cardiology

    Article Title: Genistein inhibits TNF-α-induced endothelial inflammation through the protein kinase pathway A and improves vascular inflammation in C57BL/6 mice

    doi: 10.1016/j.ijcard.2013.03.035

    Figure Lengend Snippet: Immunohistochemical staining for F4/80-positive monocytes-derived macrophages and adhesion molecule VCAM-1 in aortic cross-sections Representative photomicrographs of immunohistochemical staining for F4/80-positive monocytes-derived macrophages (Fig. 6A) and VCAM-1 (Fig. 6B). Quantitative analysis of F4/80 (Fig. 6C) and VCAM-1 (Fig. 6D). Arrows indicate typical positive stained regions and original magnification is 40×. T, TNF-α; T +G, TNF-α + genistein; Data are expressed as mean ± SEM, n=5, *, p

    Article Snippet: The adhesion molecules such as ICAM-1, VCAM-1, and E-selectin have been suggested to be atherosclerotic inflammatory markers [ , ].

    Techniques: Immunohistochemistry, Staining, Derivative Assay

    VCAM-1 changes for massage group and control group. * P

    Journal: Evidence-based Complementary and Alternative Medicine : eCAM

    Article Title: Effects of Swedish Massage Therapy on Blood Pressure, Heart Rate, and Inflammatory Markers in Hypertensive Women

    doi: 10.1155/2013/171852

    Figure Lengend Snippet: VCAM-1 changes for massage group and control group. * P

    Article Snippet: This is supported by the studies of Ando et al., (1994) [ ], Korenaga et al. (1997)[ ], and Helmlinger et al. (1995)[ ] which showed decrease in the production of VCAM-1 at physiological shear stress of > 15 dyne/cm2 and an increase in the production of VCAM-1 at shear stress of ±0–4 dyne/cm2 .

    Techniques:

    Time course of ICAM-1 and VCAM-1 expression in the rat AVM model. Xenolight-750 probe binding was examined over a period of 84 days for ICAM-1 (A) and VCAM-1 (B) in control and irradiated (15 Gy) animals (n = 7 per group). Raw MFI (mean fluorescence intensity) was normalized to day 1 in matched animals to reduce inter-animal variation. No significant differences were detected at any time between irradiated and control AVMs. Ex vivo image analysis of Xenolight 750-ICAM-1 (C) and Xenolight 750-VCAM-1 (D) probe binding in excised tissue: common carotid artery (CCA), external jugular vein (EJV) (n = 7).

    Journal: PLoS ONE

    Article Title: In vivo imaging of endothelial cell adhesion molecule expression after radiosurgery in an animal model of arteriovenous malformation

    doi: 10.1371/journal.pone.0185393

    Figure Lengend Snippet: Time course of ICAM-1 and VCAM-1 expression in the rat AVM model. Xenolight-750 probe binding was examined over a period of 84 days for ICAM-1 (A) and VCAM-1 (B) in control and irradiated (15 Gy) animals (n = 7 per group). Raw MFI (mean fluorescence intensity) was normalized to day 1 in matched animals to reduce inter-animal variation. No significant differences were detected at any time between irradiated and control AVMs. Ex vivo image analysis of Xenolight 750-ICAM-1 (C) and Xenolight 750-VCAM-1 (D) probe binding in excised tissue: common carotid artery (CCA), external jugular vein (EJV) (n = 7).

    Article Snippet: ELISA kits for ICAM-1 (BEK-2024-1P) and VCAM-1 (BEK-2107-1P; Biosensis, SA, Australia) were used according to the manufacturer’s instructions.

    Techniques: Expressing, Binding Assay, Irradiation, Fluorescence, Ex Vivo

    ELISA and immunocytochemical analysis of ICAM-1 and VCAM-1 expression in irradiated bEnd.3 cells. ELISA analysis of surface ICAM-1 (A) and VCAM-1 (D) expression in irradiated bEnd.3 cells normalized to Janus green absorbance to account for changes in cell number. Immunocytochemistry was performed on bEnd.3 cells using CF555-conjugated ICAM-1 or CF640-conjugated VCAM-1 antibodies (red). Staining was quantitated as integrated density using Image J (arbitrary units) for ICAM-1 (B) and VCAM-1 (E). Representative images are shown at 120 h (ICAM-1) (C) or 72 h (VCAM-1) (F) post-radiation at doses of 0–25 Gy. Isotype controls for ICAM-1 (IgG1-CF555) and VCAM-1 (IgG2b-CF640) showed no staining (representative images shown at 25 Gy, 72h). Cells were counterstained with DAPI to visualize nuclei (blue). All images were acquired at a magnification of 200× (scale bar = 100 μm). Values are mean ± SEM, n = 3 for each group.

    Journal: PLoS ONE

    Article Title: In vivo imaging of endothelial cell adhesion molecule expression after radiosurgery in an animal model of arteriovenous malformation

    doi: 10.1371/journal.pone.0185393

    Figure Lengend Snippet: ELISA and immunocytochemical analysis of ICAM-1 and VCAM-1 expression in irradiated bEnd.3 cells. ELISA analysis of surface ICAM-1 (A) and VCAM-1 (D) expression in irradiated bEnd.3 cells normalized to Janus green absorbance to account for changes in cell number. Immunocytochemistry was performed on bEnd.3 cells using CF555-conjugated ICAM-1 or CF640-conjugated VCAM-1 antibodies (red). Staining was quantitated as integrated density using Image J (arbitrary units) for ICAM-1 (B) and VCAM-1 (E). Representative images are shown at 120 h (ICAM-1) (C) or 72 h (VCAM-1) (F) post-radiation at doses of 0–25 Gy. Isotype controls for ICAM-1 (IgG1-CF555) and VCAM-1 (IgG2b-CF640) showed no staining (representative images shown at 25 Gy, 72h). Cells were counterstained with DAPI to visualize nuclei (blue). All images were acquired at a magnification of 200× (scale bar = 100 μm). Values are mean ± SEM, n = 3 for each group.

    Article Snippet: ELISA kits for ICAM-1 (BEK-2024-1P) and VCAM-1 (BEK-2107-1P; Biosensis, SA, Australia) were used according to the manufacturer’s instructions.

    Techniques: Enzyme-linked Immunosorbent Assay, Expressing, Irradiation, Immunocytochemistry, Staining

    In vivo near-infrared fluorescence imaging of Xenolight 750 probes in the rat AVM model. Rats with an AVM creation were sham treated or irradiated with a 15 Gy marginal dose to the AVM region by Gamma Knife and imaging performed 12 h after conjugate dye injection (25 μg/kg). Representative montages of x-ray (left), fluorescent (centre) and merged (right) images after injection of Xenolight 750 probes: (A) Xenolight-750 isotype control in irradiated animal; (B) Xenolight 750-ICAM-1 probe and; (C) Xenolight 750-VCAM-1 probe, at day 21 after sham (top panels) or radiation (bottom panels). Image J quantitation of fluorescence at day 21 post-irradiation or sham with Xenolight 750-ICAM-1 (D) or Xenolight 750-VCAM-1 (E) probes and Xenolight-750 isotype control probe.

    Journal: PLoS ONE

    Article Title: In vivo imaging of endothelial cell adhesion molecule expression after radiosurgery in an animal model of arteriovenous malformation

    doi: 10.1371/journal.pone.0185393

    Figure Lengend Snippet: In vivo near-infrared fluorescence imaging of Xenolight 750 probes in the rat AVM model. Rats with an AVM creation were sham treated or irradiated with a 15 Gy marginal dose to the AVM region by Gamma Knife and imaging performed 12 h after conjugate dye injection (25 μg/kg). Representative montages of x-ray (left), fluorescent (centre) and merged (right) images after injection of Xenolight 750 probes: (A) Xenolight-750 isotype control in irradiated animal; (B) Xenolight 750-ICAM-1 probe and; (C) Xenolight 750-VCAM-1 probe, at day 21 after sham (top panels) or radiation (bottom panels). Image J quantitation of fluorescence at day 21 post-irradiation or sham with Xenolight 750-ICAM-1 (D) or Xenolight 750-VCAM-1 (E) probes and Xenolight-750 isotype control probe.

    Article Snippet: ELISA kits for ICAM-1 (BEK-2024-1P) and VCAM-1 (BEK-2107-1P; Biosensis, SA, Australia) were used according to the manufacturer’s instructions.

    Techniques: In Vivo, Fluorescence, Imaging, Irradiation, Injection, Quantitation Assay

    Quantitative real-time PCR and western analysis of ICAM-1 and VCAM-1 expression in irradiated bEnd.3 cells. qRT-PCR analysis of ICAM-1 (A) and VCAM-1 (E) gene expression, n = 4 independent experiments. Representative western blots (time course) of ICAM-1 (B) and VCAM-1 (F) protein at 25 Gy. Representative western blots (dose response) of ICAM-1 expression (120 h) (C) and VCAM-1 expression (72 h) (G) post-irradiation. ICAM-1 (D) and VCAM-1 (H) protein expression quantitated using Image J, n = 4. Values are mean ± SEM. Data were normalized to GAPDH (westerns) or HPRT (qRT-PCR).

    Journal: PLoS ONE

    Article Title: In vivo imaging of endothelial cell adhesion molecule expression after radiosurgery in an animal model of arteriovenous malformation

    doi: 10.1371/journal.pone.0185393

    Figure Lengend Snippet: Quantitative real-time PCR and western analysis of ICAM-1 and VCAM-1 expression in irradiated bEnd.3 cells. qRT-PCR analysis of ICAM-1 (A) and VCAM-1 (E) gene expression, n = 4 independent experiments. Representative western blots (time course) of ICAM-1 (B) and VCAM-1 (F) protein at 25 Gy. Representative western blots (dose response) of ICAM-1 expression (120 h) (C) and VCAM-1 expression (72 h) (G) post-irradiation. ICAM-1 (D) and VCAM-1 (H) protein expression quantitated using Image J, n = 4. Values are mean ± SEM. Data were normalized to GAPDH (westerns) or HPRT (qRT-PCR).

    Article Snippet: ELISA kits for ICAM-1 (BEK-2024-1P) and VCAM-1 (BEK-2107-1P; Biosensis, SA, Australia) were used according to the manufacturer’s instructions.

    Techniques: Real-time Polymerase Chain Reaction, Western Blot, Expressing, Irradiation, Quantitative RT-PCR

    Summary of longitudinal changes in molecular US imaging measurements using either a P-selectin ( a ) or VCAM-1 ( b ) targeted MB contrast agent. Bar graphs detail differences between ischemic and control kidney data. (* p

    Journal: Molecular imaging and biology : MIB : the official publication of the Academy of Molecular Imaging

    Article Title: Molecular Ultrasound Imaging of Tissue Inflammation Using an Animal Model of Acute Kidney Injury

    doi: 10.1007/s11307-015-0860-6

    Figure Lengend Snippet: Summary of longitudinal changes in molecular US imaging measurements using either a P-selectin ( a ) or VCAM-1 ( b ) targeted MB contrast agent. Bar graphs detail differences between ischemic and control kidney data. (* p

    Article Snippet: Prior to image acquisition, each animal was slowly injected in the tail vein with either a 0.1 mL bolus of P-selectin (Visistar P-selectin; Targeson, San Diego, CA) or VCAM-1 (Visistar VCAM-1; Targeson) targeted MBs.

    Techniques: Imaging

    Representative histology images of renal tissue after acute kidney injury indicating that ( a ) P-selection expression was localized in the glomerulus tufts (G) and intrarenal vasculature (V) whereas ( b ) VCAM-1 upregulation was localized to the peritubular

    Journal: Molecular imaging and biology : MIB : the official publication of the Academy of Molecular Imaging

    Article Title: Molecular Ultrasound Imaging of Tissue Inflammation Using an Animal Model of Acute Kidney Injury

    doi: 10.1007/s11307-015-0860-6

    Figure Lengend Snippet: Representative histology images of renal tissue after acute kidney injury indicating that ( a ) P-selection expression was localized in the glomerulus tufts (G) and intrarenal vasculature (V) whereas ( b ) VCAM-1 upregulation was localized to the peritubular

    Article Snippet: Prior to image acquisition, each animal was slowly injected in the tail vein with either a 0.1 mL bolus of P-selectin (Visistar P-selectin; Targeson, San Diego, CA) or VCAM-1 (Visistar VCAM-1; Targeson) targeted MBs.

    Techniques: Selection, Expressing

    EC–substratum interaction influences NF-κB-dependent expression of adhesion molecules. Real-time PCR ΔΔCt of (A) VCAM-1 and (B) ICAM in MEEC (opened histogram) and EC-TCPS (closed histogram) stimulated for 6 h with 5 ng/ml TNF-α. Flow cytometry analysis of (C) VCAM and (D) ICAM expression on embedded EC (solid line), TCPS (dashed line), and TCPS with SC514 100 µM (dotted line) following 24-h stimulation with TNF-α. ≠ p

    Journal: Cell transplantation

    Article Title: NF-κB Activity in Endothelial Cells Is Modulated by Cell Substratum Interactions and Influences Chemokine-Mediated Adhesion of Natural Killer Cells

    doi: 10.3727/096368909788534979

    Figure Lengend Snippet: EC–substratum interaction influences NF-κB-dependent expression of adhesion molecules. Real-time PCR ΔΔCt of (A) VCAM-1 and (B) ICAM in MEEC (opened histogram) and EC-TCPS (closed histogram) stimulated for 6 h with 5 ng/ml TNF-α. Flow cytometry analysis of (C) VCAM and (D) ICAM expression on embedded EC (solid line), TCPS (dashed line), and TCPS with SC514 100 µM (dotted line) following 24-h stimulation with TNF-α. ≠ p

    Article Snippet: EC monolayers (incubated in 1 mmol PBS/EDTA for 5 min) or surface-adherent EC (digested with collagenase type I; Worthington Biochemical, NJ) were harvested, cell suspensions washed, and 3 × 105 EC fixed in 4% paraformaldehyde for 10 min. EC were treated with saponin (0.1% saponin, 0.05% NaN3 ) for intracellular CX3 CL1 staining (clone 51637, R & D Systems, Minneapolis, MN) or left untreated for VCAM-1 (clone 1.G11B1; Research Diagnostics, Flanders, NJ) and ICAM-1 surface staining (clone 15.2; Pharmingen, San Diego, CA) for 45 min at 4°C.

    Techniques: Expressing, Real-time Polymerase Chain Reaction, Flow Cytometry, Cytometry

    Mechanisms of lymphocyte infiltration and localization of virus in pediatric lymphocytic interstitial pneumonia. A: Longitudinal section of a vein showing VCAM-1 expression and associated mononuclear cell infiltrates. IP for VCAM-1 (clone 51–10C9, PharMingen); magnification, ×100. The inset shows a transverse section of a venule and demonstrates the uniform expression of endothelial VCAM-1. IP for VCAM-1; magnification, ×400. B: Perivenular aggregation of CD8 + T lymphocytes in areas of interstitial collapse. IP for CD8 (clone C8/144B, DAKO); magnification, ×400. C: Localization of cells harboring HIV mRNA ( arrows ) within a thin-walled vein. 125 I ISH for HIV-1 mRNA; magnification, ×100. D: Expression of CMV early gene RNA in cytomegaloid cells ( arrows ) adjacent to a vein and in a region of interstitial collapse and lymphocyte infiltration. ISH αDIG-AP for EBV EBER-1 RNA; magnification, ×400. E: Expression of KSHV latent gene transcripts in cells associated with follicular aggregation. ISH αDIG-AP for KSHV T0.7 RNA; magnification, ×400.

    Journal: The American Journal of Pathology

    Article Title: Pediatric AIDS-Associated Lymphocytic Interstitial Pneumonia and Pulmonary Arterio-Occlusive Disease

    doi:

    Figure Lengend Snippet: Mechanisms of lymphocyte infiltration and localization of virus in pediatric lymphocytic interstitial pneumonia. A: Longitudinal section of a vein showing VCAM-1 expression and associated mononuclear cell infiltrates. IP for VCAM-1 (clone 51–10C9, PharMingen); magnification, ×100. The inset shows a transverse section of a venule and demonstrates the uniform expression of endothelial VCAM-1. IP for VCAM-1; magnification, ×400. B: Perivenular aggregation of CD8 + T lymphocytes in areas of interstitial collapse. IP for CD8 (clone C8/144B, DAKO); magnification, ×400. C: Localization of cells harboring HIV mRNA ( arrows ) within a thin-walled vein. 125 I ISH for HIV-1 mRNA; magnification, ×100. D: Expression of CMV early gene RNA in cytomegaloid cells ( arrows ) adjacent to a vein and in a region of interstitial collapse and lymphocyte infiltration. ISH αDIG-AP for EBV EBER-1 RNA; magnification, ×400. E: Expression of KSHV latent gene transcripts in cells associated with follicular aggregation. ISH αDIG-AP for KSHV T0.7 RNA; magnification, ×400.

    Article Snippet: When these tissues and cells were pretreated with MAbs to VCAM-1 or VLA-4α4/β1 , respectively, the MBC dropped to < 5.

    Techniques: Expressing, In Situ Hybridization

    Outgrowth endothelial cell morphology and phenotype. Outgrowth cells have typical endothelial morphology ( a ). The remaining parts ( b – e ) show, as labeled at the bottom of each, constitutive and activated phenotype by immunofluorescent staining. Outgrowth endothelial cells incorporated acetylated LDL and were positive for vWF. They were negative for VCAM-1 but expressed it upon stimulation.

    Journal: Journal of Clinical Investigation

    Article Title: Origins of circulating endothelial cells and endothelial outgrowth from blood

    doi:

    Figure Lengend Snippet: Outgrowth endothelial cell morphology and phenotype. Outgrowth cells have typical endothelial morphology ( a ). The remaining parts ( b – e ) show, as labeled at the bottom of each, constitutive and activated phenotype by immunofluorescent staining. Outgrowth endothelial cells incorporated acetylated LDL and were positive for vWF. They were negative for VCAM-1 but expressed it upon stimulation.

    Article Snippet: To deliberately activate endothelial cells, they were incubated at 37°C for 4 hours with either 10 ng/mL of IL-1 for stimulation of VCAM-1 and ICAM-1, or 10 ng/mL of LPS for tissue factor stimulation.

    Techniques: Labeling, Staining

    Results of gene expression by Semi-quantitative RT-PCR. Representative gel images of E-sel, ICAM-1, VCAM-1 and MCP- 1 mRNA expression of the second portion of rabbit aortas. β-actin was used as an internal control and PCR products were visualized by ethidium bromide staining. The data in the bar graph are quantified ratios of the signal for E-sel, ICAM-1, VCAM-1 and MCP-1 to that for β-actin with the control samples set at 100%. Results are expressed as mean± SD (n = 6). ** p

    Journal: Lipids in Health and Disease

    Article Title: The anti-inflammatory effect of kaempferol on early atherosclerosis in high cholesterol fed rabbits

    doi: 10.1186/1476-511X-12-115

    Figure Lengend Snippet: Results of gene expression by Semi-quantitative RT-PCR. Representative gel images of E-sel, ICAM-1, VCAM-1 and MCP- 1 mRNA expression of the second portion of rabbit aortas. β-actin was used as an internal control and PCR products were visualized by ethidium bromide staining. The data in the bar graph are quantified ratios of the signal for E-sel, ICAM-1, VCAM-1 and MCP-1 to that for β-actin with the control samples set at 100%. Results are expressed as mean± SD (n = 6). ** p

    Article Snippet: In our study, mRNA and protein expressions of E-sel, ICAM-1, VCAM-1 and MCP-1 in aorta of rabbits were all up-regulated by a high-cholesterol diet and expressions of those three molecules were all down-regulated by kaempferol (30 mg/kg and 150 mg/kg).

    Techniques: Expressing, Quantitative RT-PCR, Polymerase Chain Reaction, Staining

    Results of immunohistochemical study. a . Representative examples of immunohistochemical staining of E-sel (A-E), ICAM-1 (F-J), VCAM-1 (K-O) and MCP-1 (P-T) in the first portion of rabbit aortas (amplification ×400). b . Immunohistochemical score (IHS) of rabbit aorta sections. Control: rabbits fed on normal diets; Model: rabbits fed on high-cholesterol diets; Fenofibrate: rabbits received high-cholesterol diets plus fenofibrate (12 mg/kg); Kaempferol-H: rabbits received high-cholesterol diets plus kaempferol (150 mg/kg); Kaempferol-L: rabbits received high-cholesterol diets plus kaempferol (30 mg/kg). Each bar represents mean± SD (n = 6). * P

    Journal: Lipids in Health and Disease

    Article Title: The anti-inflammatory effect of kaempferol on early atherosclerosis in high cholesterol fed rabbits

    doi: 10.1186/1476-511X-12-115

    Figure Lengend Snippet: Results of immunohistochemical study. a . Representative examples of immunohistochemical staining of E-sel (A-E), ICAM-1 (F-J), VCAM-1 (K-O) and MCP-1 (P-T) in the first portion of rabbit aortas (amplification ×400). b . Immunohistochemical score (IHS) of rabbit aorta sections. Control: rabbits fed on normal diets; Model: rabbits fed on high-cholesterol diets; Fenofibrate: rabbits received high-cholesterol diets plus fenofibrate (12 mg/kg); Kaempferol-H: rabbits received high-cholesterol diets plus kaempferol (150 mg/kg); Kaempferol-L: rabbits received high-cholesterol diets plus kaempferol (30 mg/kg). Each bar represents mean± SD (n = 6). * P

    Article Snippet: In our study, mRNA and protein expressions of E-sel, ICAM-1, VCAM-1 and MCP-1 in aorta of rabbits were all up-regulated by a high-cholesterol diet and expressions of those three molecules were all down-regulated by kaempferol (30 mg/kg and 150 mg/kg).

    Techniques: Immunohistochemistry, Staining, Amplification

    Bilirubin suppresses cellular  ROS  generation following activation of  VCAM ‐1 or  ICAM ‐1.  TNF ‐α‐activated  HUVEC  monolayers were incubated with anti‐ VCAM ‐1 (left panels, α VCAM ‐1; 10 μg/mL) or anti‐ ICAM ‐1 (right panels, α ICAM ‐1; 10 μg/mL) for 30 minutes and then loaded with dihydrorhodamine. Adhesion molecule activation was triggered by the addition of a cross‐linking antibody and  ROS  generation quantified by confocal microscopy. Upper panels display representative time‐lapse images of nonstimulated and antibody‐activated cells treated with 20 μmol/L of bilirubin ( BR ) or the bilirubin vehicle (Veh). Scale bars represent 100 μm. Lower panels plot the time‐dependent changes in fluorescence intensity following  VCAM ‐1 (left panel) or  ICAM ‐1 (right panel) activation (squares), in the presence (white symbols) or absence (black symbols) of bilirubin. Cells that were not treated with cross‐linking antibodies (nonstimulated; circles) serve as control, with curves reflecting mean fluorescence intensity (±SEM) expressed relative to maximal activation at 60 minutes (n=3 sets of experiments). * P

    Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

    Article Title: Bilirubin Prevents Atherosclerotic Lesion Formation in Low‐Density Lipoprotein Receptor‐Deficient Mice by Inhibiting Endothelial VCAM‐1 and ICAM‐1 Signaling

    doi: 10.1161/JAHA.116.004820

    Figure Lengend Snippet: Bilirubin suppresses cellular ROS generation following activation of VCAM ‐1 or ICAM ‐1. TNF ‐α‐activated HUVEC monolayers were incubated with anti‐ VCAM ‐1 (left panels, α VCAM ‐1; 10 μg/mL) or anti‐ ICAM ‐1 (right panels, α ICAM ‐1; 10 μg/mL) for 30 minutes and then loaded with dihydrorhodamine. Adhesion molecule activation was triggered by the addition of a cross‐linking antibody and ROS generation quantified by confocal microscopy. Upper panels display representative time‐lapse images of nonstimulated and antibody‐activated cells treated with 20 μmol/L of bilirubin ( BR ) or the bilirubin vehicle (Veh). Scale bars represent 100 μm. Lower panels plot the time‐dependent changes in fluorescence intensity following VCAM ‐1 (left panel) or ICAM ‐1 (right panel) activation (squares), in the presence (white symbols) or absence (black symbols) of bilirubin. Cells that were not treated with cross‐linking antibodies (nonstimulated; circles) serve as control, with curves reflecting mean fluorescence intensity (±SEM) expressed relative to maximal activation at 60 minutes (n=3 sets of experiments). * P

    Article Snippet: Our demonstration that bilirubin attenuates ROS generation by activated HUVECs in response to VCAM‐1 or ICAM‐1 cross‐linking and inhibits H2 O2 production by isolated XO enzyme supports this hypothesis.

    Techniques: Activation Assay, Incubation, Confocal Microscopy, Fluorescence

    Bilirubin does not alter adhesion molecule expression or monocyte binding to  HUVECs .  HUVEC  monolayers were incubated with or without  TNF ‐α (5 ng/mL) in the presence of bilirubin ( BR ; 20 μmol/L) or the bilirubin vehicle (Veh). A,  mRNA  levels for  ICAM ‐1,  VCAM ‐1,  PECAM ‐1, E‐Selectin, and P‐Selectin at baseline and at 4 hours (n=4 sets of experiments). The right lower panel depicts the results of an adhesion assay measuring the binding of CellTrace Far Red–labeled  THP ‐1 cells to  HUVEC  monolayers incubated with or without  TNF ‐α for 24 hours. Bars reflect mean fluorescence intensity (±SEM) relative to non‐ TNF ‐α‐activated  HUVECs  (n=4 sets of experiments). B, Representative immunoblots for E‐Selectin,  ICAM ‐1, and  VCAM ‐1, which are quantified (C) as described in Figure   1  (n=4 sets of experiments). * P

    Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

    Article Title: Bilirubin Prevents Atherosclerotic Lesion Formation in Low‐Density Lipoprotein Receptor‐Deficient Mice by Inhibiting Endothelial VCAM‐1 and ICAM‐1 Signaling

    doi: 10.1161/JAHA.116.004820

    Figure Lengend Snippet: Bilirubin does not alter adhesion molecule expression or monocyte binding to HUVECs . HUVEC monolayers were incubated with or without TNF ‐α (5 ng/mL) in the presence of bilirubin ( BR ; 20 μmol/L) or the bilirubin vehicle (Veh). A, mRNA levels for ICAM ‐1, VCAM ‐1, PECAM ‐1, E‐Selectin, and P‐Selectin at baseline and at 4 hours (n=4 sets of experiments). The right lower panel depicts the results of an adhesion assay measuring the binding of CellTrace Far Red–labeled THP ‐1 cells to HUVEC monolayers incubated with or without TNF ‐α for 24 hours. Bars reflect mean fluorescence intensity (±SEM) relative to non‐ TNF ‐α‐activated HUVECs (n=4 sets of experiments). B, Representative immunoblots for E‐Selectin, ICAM ‐1, and VCAM ‐1, which are quantified (C) as described in Figure  1 (n=4 sets of experiments). * P

    Article Snippet: Our demonstration that bilirubin attenuates ROS generation by activated HUVECs in response to VCAM‐1 or ICAM‐1 cross‐linking and inhibits H2 O2 production by isolated XO enzyme supports this hypothesis.

    Techniques: Expressing, Binding Assay, Incubation, Cell Adhesion Assay, Labeling, Fluorescence, Western Blot

    Bilirubin does not alter VCAM ‐1 or ICAM ‐1 expression in Ldlr −/− mice. A and B, Representative photomicrographs of sections of aortic root stained with immunofluorescent antibodies against VCAM ‐1 (green; A) or ICAM ‐1 (cyan; B). Data are presented and analyzed as described in Figure 9 . DAPI indicates 4',6‐diamidino‐2‐phenylindole; ICAM‐1, intercellular adhesion molecule 1; VCAM‐1, vascular cell adhesion molecule 1.

    Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

    Article Title: Bilirubin Prevents Atherosclerotic Lesion Formation in Low‐Density Lipoprotein Receptor‐Deficient Mice by Inhibiting Endothelial VCAM‐1 and ICAM‐1 Signaling

    doi: 10.1161/JAHA.116.004820

    Figure Lengend Snippet: Bilirubin does not alter VCAM ‐1 or ICAM ‐1 expression in Ldlr −/− mice. A and B, Representative photomicrographs of sections of aortic root stained with immunofluorescent antibodies against VCAM ‐1 (green; A) or ICAM ‐1 (cyan; B). Data are presented and analyzed as described in Figure 9 . DAPI indicates 4',6‐diamidino‐2‐phenylindole; ICAM‐1, intercellular adhesion molecule 1; VCAM‐1, vascular cell adhesion molecule 1.

    Article Snippet: Our demonstration that bilirubin attenuates ROS generation by activated HUVECs in response to VCAM‐1 or ICAM‐1 cross‐linking and inhibits H2 O2 production by isolated XO enzyme supports this hypothesis.

    Techniques: Expressing, Mouse Assay, Staining

    Nox and  XO  inhibitors and antibodies against  VCAM ‐1 and  ICAM ‐1 recapitulate the effect of bilirubin on endothelial  ROS  generation and monocyte transmigration.  ROS  production by  TNF ‐α‐stimulated  HUVEC  monolayers was assessed by monitoring dihydrorhodamine fluorescence following activation of  VCAM ‐1 (α VCAM ‐1) or  ICAM ‐1 (α ICAM ‐1), as described in Figure   5 . A and B, Time‐dependent changes in fluorescence intensity following  VCAM ‐1 (A) or  ICAM ‐1 (B) activation (squares), in the absence (black symbols) or presence of 10 μmol/L of  ML 171 (white symbols), or 40 μmol/L of allopurinol ( AP ; gray symbols). Curves reflect mean fluorescence intensity (±SEM) expressed relative to maximal activation at 60 minutes (n=3 sets of experiments). C, Compares the effect of the  DMSO  vehicle (Veh), 40 μmol/L of  AP , and/or 10 μmol/L of  ML 171 on  THP ‐1 cell migration across  HUVEC  monolayers, as described in Figure   2  (n=4 sets of experiments). D and E, Results of analogous studies examining  THP ‐1 migration in the presence or absence of antibodies against  ICAM ‐1 ( ICAM ; 10 μg/mL),  VCAM ‐1 ( VCAM ; 10 μg/mL), β 2  (5 μg/mL), and/or α 4  (20 μg/mL). F, Lineweaver–Burk plot of H 2 O 2  produced by isolated XO in the presence of 50 μmol/L of bilirubin ( BR ; diamonds; K i =3.4 μmol/L), 30 μmol/L of the competitive inhibitor, AP (triangles; K i =6.7 μmol/L), 30 μmol/L of the noncompetitive inhibitor, 2‐chloro‐6(methylamino) purine ( CMAP ; squares; K i =4.7 μmol/L), or the  DMSO  vehicle (circles). Data reflect the mean (±SEM) of 3 sets of experiments. * P

    Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

    Article Title: Bilirubin Prevents Atherosclerotic Lesion Formation in Low‐Density Lipoprotein Receptor‐Deficient Mice by Inhibiting Endothelial VCAM‐1 and ICAM‐1 Signaling

    doi: 10.1161/JAHA.116.004820

    Figure Lengend Snippet: Nox and XO inhibitors and antibodies against VCAM ‐1 and ICAM ‐1 recapitulate the effect of bilirubin on endothelial ROS generation and monocyte transmigration. ROS production by TNF ‐α‐stimulated HUVEC monolayers was assessed by monitoring dihydrorhodamine fluorescence following activation of VCAM ‐1 (α VCAM ‐1) or ICAM ‐1 (α ICAM ‐1), as described in Figure  5 . A and B, Time‐dependent changes in fluorescence intensity following VCAM ‐1 (A) or ICAM ‐1 (B) activation (squares), in the absence (black symbols) or presence of 10 μmol/L of ML 171 (white symbols), or 40 μmol/L of allopurinol ( AP ; gray symbols). Curves reflect mean fluorescence intensity (±SEM) expressed relative to maximal activation at 60 minutes (n=3 sets of experiments). C, Compares the effect of the DMSO vehicle (Veh), 40 μmol/L of AP , and/or 10 μmol/L of ML 171 on THP ‐1 cell migration across HUVEC monolayers, as described in Figure  2 (n=4 sets of experiments). D and E, Results of analogous studies examining THP ‐1 migration in the presence or absence of antibodies against ICAM ‐1 ( ICAM ; 10 μg/mL), VCAM ‐1 ( VCAM ; 10 μg/mL), β 2 (5 μg/mL), and/or α 4 (20 μg/mL). F, Lineweaver–Burk plot of H 2 O 2 produced by isolated XO in the presence of 50 μmol/L of bilirubin ( BR ; diamonds; K i =3.4 μmol/L), 30 μmol/L of the competitive inhibitor, AP (triangles; K i =6.7 μmol/L), 30 μmol/L of the noncompetitive inhibitor, 2‐chloro‐6(methylamino) purine ( CMAP ; squares; K i =4.7 μmol/L), or the DMSO vehicle (circles). Data reflect the mean (±SEM) of 3 sets of experiments. * P

    Article Snippet: Our demonstration that bilirubin attenuates ROS generation by activated HUVECs in response to VCAM‐1 or ICAM‐1 cross‐linking and inhibits H2 O2 production by isolated XO enzyme supports this hypothesis.

    Techniques: Transmigration Assay, Fluorescence, Activation Assay, Migration, Produced, Isolation

    Proposed mechanism of bilirubin modulation of VCAM ‐1‐ and ICAM ‐1‐dependent monocyte migration. Ligation of VCAM ‐1 and ICAM ‐1 with their corresponding integrins, α 4 β 1 /α 4 β 7 and α L β 2 , leads to Rac‐1‐ and calcium (Ca 2+ )‐dependent activation of NADPH oxidase (Nox) and xanthine oxidase ( XO ). These enzymes generate the reactive oxygen species ( ROS ), superoxide (O 2 ˙ − ) and hydrogen peroxide (H 2 O 2 ), that comprise a signaling cascade, which leads to activation of matrix metalloproteinases ( MMP )‐2 and ‐9 and disruption of endothelial tight junctions. Bilirubin, a potent antioxidant that undergoes intracellular redox cycling (dashed lines) through action of biliverdin reductase ( BVR ), scavenges Nox‐ and XO ‐derived ROS , thereby inhibiting leukocyte migration. ICAM‐1 indicates intercellular adhesion molecule 1; Rac1, Ras‐related C3 botulinum toxin substrate 1; VCAM‐1, vascular cell adhesion molecule 1.

    Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

    Article Title: Bilirubin Prevents Atherosclerotic Lesion Formation in Low‐Density Lipoprotein Receptor‐Deficient Mice by Inhibiting Endothelial VCAM‐1 and ICAM‐1 Signaling

    doi: 10.1161/JAHA.116.004820

    Figure Lengend Snippet: Proposed mechanism of bilirubin modulation of VCAM ‐1‐ and ICAM ‐1‐dependent monocyte migration. Ligation of VCAM ‐1 and ICAM ‐1 with their corresponding integrins, α 4 β 1 /α 4 β 7 and α L β 2 , leads to Rac‐1‐ and calcium (Ca 2+ )‐dependent activation of NADPH oxidase (Nox) and xanthine oxidase ( XO ). These enzymes generate the reactive oxygen species ( ROS ), superoxide (O 2 ˙ − ) and hydrogen peroxide (H 2 O 2 ), that comprise a signaling cascade, which leads to activation of matrix metalloproteinases ( MMP )‐2 and ‐9 and disruption of endothelial tight junctions. Bilirubin, a potent antioxidant that undergoes intracellular redox cycling (dashed lines) through action of biliverdin reductase ( BVR ), scavenges Nox‐ and XO ‐derived ROS , thereby inhibiting leukocyte migration. ICAM‐1 indicates intercellular adhesion molecule 1; Rac1, Ras‐related C3 botulinum toxin substrate 1; VCAM‐1, vascular cell adhesion molecule 1.

    Article Snippet: Our demonstration that bilirubin attenuates ROS generation by activated HUVECs in response to VCAM‐1 or ICAM‐1 cross‐linking and inhibits H2 O2 production by isolated XO enzyme supports this hypothesis.

    Techniques: Migration, Ligation, Activation Assay, Derivative Assay

    Time course for  TNF ‐α‐induced expression of adhesion molecules by  HUVECs .  HUVEC  monolayers were incubated in the presence of 5 ng/mL of  TNF ‐α (TNF; squares) or the  TNF  vehicle (Veh; circles), and expression of  VCAM ‐1,  ICAM ‐1, E‐Selectin, P‐Selectin, and  PECAM ‐1 was determined at the indicated time points by  qRT ‐ PCR  and western blotting. A, Time‐dependent changes in  mRNA  for E‐Selectin (black symbols) and P‐Selectin (white symbols), while (B) displays the results obtained for  VCAM ‐1 (gray symbols),  ICAM ‐1 (black symbols), and  PECAM ‐1 (white symbols). Data reflect  mRNA  levels (±SEM) relative to untreated cells (n=4 separate sets of experiments). C, Representative immunoblots for E‐Selectin,  ICAM ‐1, and  VCAM ‐1, with graphs (D) quantifying expression at the indicated time points relative to unstimulated cells at time 0 (Con) and corrected for  GAPDH  (n=3 sets of experiments). * P

    Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

    Article Title: Bilirubin Prevents Atherosclerotic Lesion Formation in Low‐Density Lipoprotein Receptor‐Deficient Mice by Inhibiting Endothelial VCAM‐1 and ICAM‐1 Signaling

    doi: 10.1161/JAHA.116.004820

    Figure Lengend Snippet: Time course for TNF ‐α‐induced expression of adhesion molecules by HUVECs . HUVEC monolayers were incubated in the presence of 5 ng/mL of TNF ‐α (TNF; squares) or the TNF vehicle (Veh; circles), and expression of VCAM ‐1, ICAM ‐1, E‐Selectin, P‐Selectin, and PECAM ‐1 was determined at the indicated time points by qRT ‐ PCR and western blotting. A, Time‐dependent changes in mRNA for E‐Selectin (black symbols) and P‐Selectin (white symbols), while (B) displays the results obtained for VCAM ‐1 (gray symbols), ICAM ‐1 (black symbols), and PECAM ‐1 (white symbols). Data reflect mRNA levels (±SEM) relative to untreated cells (n=4 separate sets of experiments). C, Representative immunoblots for E‐Selectin, ICAM ‐1, and VCAM ‐1, with graphs (D) quantifying expression at the indicated time points relative to unstimulated cells at time 0 (Con) and corrected for GAPDH (n=3 sets of experiments). * P

    Article Snippet: Our demonstration that bilirubin attenuates ROS generation by activated HUVECs in response to VCAM‐1 or ICAM‐1 cross‐linking and inhibits H2 O2 production by isolated XO enzyme supports this hypothesis.

    Techniques: Expressing, Incubation, Quantitative RT-PCR, Western Blot

    MPIO-contrast volume reflects VCAM-1 mRNA expression in tissue. VCAM-1 mRNA expression was 12-fold higher in clamped kidneys versus sham operated kidneys (n = 5, P

    Journal: PLoS ONE

    Article Title: In Vivo Quantification of Vcam-1 Expression in Renal Ischemia Reperfusion Injury Using Non-Invasive Magnetic Resonance Molecular Imaging

    doi: 10.1371/journal.pone.0012800

    Figure Lengend Snippet: MPIO-contrast volume reflects VCAM-1 mRNA expression in tissue. VCAM-1 mRNA expression was 12-fold higher in clamped kidneys versus sham operated kidneys (n = 5, P

    Article Snippet: Distribution of VCAM-1 and microparticles of iron oxide on histology On microscopic inspection of IRI kidneys, VCAM-MPIO were adherent to the vessel wall, either singly or in small clusters, of the peritubular capillaries ( ).

    Techniques: Expressing

    3D reconstruction of segmented kidneys. A . VCAM-MPIO contrast (green) was abundant and in both the medulla and cortex of IRI kidneys (image-left) and to a lesser extent in sham-operated kidneys (image-right). B . In mice that underwent an identical protocol, there was little to no binding of the isotype IgG-MPIO in either kidney. C. Similarly, there was little to no VCAM-MPIO retention in either kidney in mice that underwent no surgery (n = 2) or D . in mice pre-treated with anti-VCAM-1 antibody prior to contrast administration.

    Journal: PLoS ONE

    Article Title: In Vivo Quantification of Vcam-1 Expression in Renal Ischemia Reperfusion Injury Using Non-Invasive Magnetic Resonance Molecular Imaging

    doi: 10.1371/journal.pone.0012800

    Figure Lengend Snippet: 3D reconstruction of segmented kidneys. A . VCAM-MPIO contrast (green) was abundant and in both the medulla and cortex of IRI kidneys (image-left) and to a lesser extent in sham-operated kidneys (image-right). B . In mice that underwent an identical protocol, there was little to no binding of the isotype IgG-MPIO in either kidney. C. Similarly, there was little to no VCAM-MPIO retention in either kidney in mice that underwent no surgery (n = 2) or D . in mice pre-treated with anti-VCAM-1 antibody prior to contrast administration.

    Article Snippet: Distribution of VCAM-1 and microparticles of iron oxide on histology On microscopic inspection of IRI kidneys, VCAM-MPIO were adherent to the vessel wall, either singly or in small clusters, of the peritubular capillaries ( ).

    Techniques: Mouse Assay, Binding Assay

    Serial in-vivo MRI at 30, 60 and 90 minutes detects VCAM-MPIO accumulation in renal ischemia-reperfusion injury (IRI). A . VCAM-MPIO caused a marked contrast effect evident as low-signal in the renal cortex and medulla of the IRI kidney (red box). A lesser degree of signal loss was also evident in the contra-lateral, sham-operated kidney (green box). B . Irrelevant isotype IgG-MPIO control showed no contrast effect in either IRI (red box) or sham-operated kidneys (green box) (n = 3). C . Pre-treatment of mice with anti-VCAM-1 antibody abolished retention of VCAM-MPIO in both IRI (red box) and sham-operated (green box) kidneys. In A–C, contrast effects peaked at 60 minutes and were sustained throughout the imaging protocol. Scale bar = 5 mm.

    Journal: PLoS ONE

    Article Title: In Vivo Quantification of Vcam-1 Expression in Renal Ischemia Reperfusion Injury Using Non-Invasive Magnetic Resonance Molecular Imaging

    doi: 10.1371/journal.pone.0012800

    Figure Lengend Snippet: Serial in-vivo MRI at 30, 60 and 90 minutes detects VCAM-MPIO accumulation in renal ischemia-reperfusion injury (IRI). A . VCAM-MPIO caused a marked contrast effect evident as low-signal in the renal cortex and medulla of the IRI kidney (red box). A lesser degree of signal loss was also evident in the contra-lateral, sham-operated kidney (green box). B . Irrelevant isotype IgG-MPIO control showed no contrast effect in either IRI (red box) or sham-operated kidneys (green box) (n = 3). C . Pre-treatment of mice with anti-VCAM-1 antibody abolished retention of VCAM-MPIO in both IRI (red box) and sham-operated (green box) kidneys. In A–C, contrast effects peaked at 60 minutes and were sustained throughout the imaging protocol. Scale bar = 5 mm.

    Article Snippet: Distribution of VCAM-1 and microparticles of iron oxide on histology On microscopic inspection of IRI kidneys, VCAM-MPIO were adherent to the vessel wall, either singly or in small clusters, of the peritubular capillaries ( ).

    Techniques: In Vivo, Magnetic Resonance Imaging, Mouse Assay, Imaging