cd31 Search Results


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  • 99
    Thermo Fisher anti cd31
    Representative figures of cellular atypia (A), <t>CD31</t> expression levels (B) and vWF expression levels (C). Inserted images are the magnified views of each figures. Rectangle images at the bottom of (B) and (C) are immunohistochemical reactivity of CD31 and vWF in normal endothelial cells in the same slides of above images. Bars=20 µ m.
    Anti Cd31, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 864 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    cd31  (Abcam)
    94
    Abcam cd31
    Endothelial phenotype changes with stromal cell co-culture. a Endothelial staining of HS27a and b HS5 co-cultured vessels shows differences in (i) <t>CD31,</t> (ii) VE-cadherin (VE-Cad), and (iii) von Willebrand Factor (vWF). c RNA expression within the co-culture shows differences in endothelial junctional expression and inflammatory marker expression. * p
    Cd31, supplied by Abcam, used in various techniques. Bioz Stars score: 94/100, based on 6840 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Becton Dickinson cd31
    Relationship between HSC-primed HECs and T1 pre-HSCs. (a) Representative FACS plots for sorting of the T1 pre-HSCs <t>(CD31</t> + CD45 - CD41 low Kit + CD201 high ) from E11.0 AGM region of mouse embryos. Red box indicates the sampling cells for scRNA-seq. (b) Violin plots showing the expression levels of indicated genes in tif-HEC (including cluster HEC and PK44), T1 pre-HSC and lateAEC. (c) t-SNE plot of the cells included in the filtered initial dataset, PK44 dataset and T1 pre-HSC dataset, with clusters mapped on it. Cluster HEC and PK44 are combined as tif-HEC. (d) PCA plot of tif-HEC and T1 pre-HSC populations. (e) Enriched terms of PC2 positive and negative genes are shown, corresponding to the properties distinguishing tif-HEC and T1 pre-HSC, respectively. (f) Heatmap showing top 20 positive and negative genes of PC2. Genes were ordered by their contributions to PC2. (g) Trajectory of AEC clusters, tif-HEC and T1 pre-HSC inferred by Mpath. Arrows indicate the development directions predicted by sampling stages. (h) Representative FACS plots for sorting of the PK44 cells from E10.0 AGM region (left) and analysis of the immunophenotypic T1 pre-HSCs (right) after cultured in vitro for 4 days. (i) Representative CD31 immunostaining on the cultures of single T1 pre-HSCs from E11.0 AGM region, showing typical morphologies regarding distinct differentiation capacities. Cell frequencies of each type are also shown. Data are from 7 independent experiments with totally 89 embryos used. Scale bars, 400 μm.
    Cd31, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 92/100, based on 8574 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Santa Cruz Biotechnology cd31
    Increased tumor hypoxia in obese mice. Tumors in mice fed a high-fat diet (HFD) have higher Ki67+ counts ( a , scalebar 200 um). <t>CD31</t> immunohistochemical analysis shows lower vessel density in tumors from HFD mice ( b , scalebar 200 uM). This is supported by fluorescence-activated cell sorting quantification ( c ). Pimonidazole (PIMO) staining in mouse fed a regular diet (RD) or a HFD demonstrate greater hypoxic areas in tumors from HFD mice ( d , scalebar 200 uM). Quantitative PCR analyses on RD and HFD tumors show upregulation of hypoxia inducible factor 1 alpha (HIF1α) targets ( e , n = 5) and faster tumor progression ( f , n = 5). Arbp, attachment region binding protein
    Cd31, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 92/100, based on 3022 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Agilent technologies cd31
    Macroscopic and microscopic features of the plugs two and eight weeks after implantation. Plugs were harvested two ( A ) and eight weeks ( B ) after subcutaneous co-transplantation of ECFC+MSPC (ratio 80∶ 20, left column), MSPC only (2 nd column), ECFC only (3 rd column), and cell-free matrix (right column). Representative macroscopic pictures of plugs in subcutaneous location (upper rows), micrographs of hematoxylin and eosin staining (HE, middle rows) and immune histochemistry combining human <t>CD31</t> colored brown with hematoxylin counter-stain (huCD31, bottom rows) are depicted. Total cell number within the plugs was 2×10 6 per 300 µL matrix (n = 3 per group and time course).
    Cd31, supplied by Agilent technologies, used in various techniques. Bioz Stars score: 94/100, based on 3427 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Becton Dickinson anti cd31
    Loss of TSP-1 promotes tumor angiogenesis and platelet infiltration. (A-C) Tumors from WT and TSP-1 null mice were sectioned and stained with <t>CD31</t> for microvessels, SMA for mature vessels, and CD42 for platelets. Quantification of staining per tumor is represented as mean±SEM ( n =4). * represents p
    Anti Cd31, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 94/100, based on 3140 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Abcam anti cd31 antibody
    Average cell populations for SVF constructs fixed after initial incubation and 48-hours of dynamic culture (n = 6). A paired t-test indicated no significant statistical difference between initial and 48-hour constructs for each of the CD90, <t>CD31,</t> and CD34 markers (P = 0.63, 0.68, and 0.46 respectively).
    Anti Cd31 Antibody, supplied by Abcam, used in various techniques. Bioz Stars score: 92/100, based on 1561 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Becton Dickinson rat anti mouse cd31
    Angiogenic response in virus-induced scars. Endothelial <t>CD31</t> expression (red) marks blood vessels, PDGFRα in green and lipid membranes in grey. (Left column, A, C, E, G, I, K) Overview, showing changes in vessel morphology and density in PDGF induced scars. (Right column, B, D, F, H, J, L) Asterisks mark big vessels with altered morphology. (M) Quantification of small vessels in the different experimental groups, normalized to the empty vector control. (N) Quantification of vessel size (area of cross-sectioned vessels) for the different experimental groups, normalized to the empty vector control. Analyses were performed on 3–4 mice per experimental group. E = empty vector; As = PDGF-A short ; Al = PDGF-A long ; B = PDGF-B; C = PDGF-C; D = PDGF-D. Scale bars are 100 μm.
    Rat Anti Mouse Cd31, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 93/100, based on 2558 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Thermo Fisher cd31
    Comparison of kappa values among pathologists for lymphovascular invasion (LVI) detection in colorectal cancers. While the average of LVI detection rate for each pathologist was 43% with hematoxylin and eosin (H E) only, 10% with <t>CD31,</t> 29% with D2-40, and 16% with ERG, the consensus reached 80% of LVI detection after a joint discussion about ERG patterns with LVI. a Interpreted by ERG
    Cd31, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 2560 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Becton Dickinson rat anti cd31
    Moesin is expressed by microglia and endothelial cells in mouse brain. A Representative immunofluorescence images of Cx3cr1 CreER-YFP -WT ( N = 4) and Cx3cr1 CreER-YFP -5xFAD ( N = 6) mouse cortex stained for microglia (GFP) and Msn. Arrow indicates microglia immunopositive for GFP and Msn. Asterisk indicates cells immunopositive for Msn only. B Representative immunofluorescence images of Cx3cr1 CreER-YFP -5xFAD ( N = 6) mouse cortex stained for amyloid-beta (Aβ), microglia (GFP), and Msn. Arrow indicates Aβ plaque as well as microglia immunopositive for GFP and Msn. C Representative immunofluorescence images of WT ( N = 3) and 5xFAD ( N = 4) mouse cortex stained for endothelial cells <t>(CD31)</t> and Msn. Arrow indicates endothelial cells immunopositive for CD31 and Msn. Asterisk indicates cells immunopostive for Msn only. D Representative immunofluorescence images of WT ( N = 3) and 5xFAD ( N = 4) mouse cortex stained for astrocytes (GFAP) and Msn. Arrow indicates cells immunopositive for Msn only. Asterisk indicates astroctyes immunopostive for GFAP only. Scale bar = 30µm.
    Rat Anti Cd31, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 94/100, based on 1675 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Cell Signaling Technology Inc cd31
    Endothelium contributes cells to aortic calcification of Ins2 Akita/+ mice (A) Aortic expression of endothelial markers <t>CD31</t> and vWF in Ins2 Akita/+ mice visualized by immunostaining. (B) Immunostaining of aortic tissues from wild type (WT) and Ins2 Akita/+ mice showed co-expression of endothelial markers CD31 (left) and vWF (right) and osteogenic markers Cbfa1 and Osterix (OSX) in the Ins2 Akita/+ mice. (C) Co-expression of CD31 and Cbfa1 in enzymatically dispersed aortic cells from WT and Ins2 Akita/+ mice , as determined by FACS. (D) Aortic expression of GFP in Tie2-Gfp tg and Ins2 Akita/+ ;Tie2-Gfp tg mice by immunostaining with anti-GFP antibodies. (E) Immunostaining of aortic tissues from Tie2-Gfp tg and Ins2 Akita/+ ;Tie2-Gfp tg mice showed co-expression of GFP with Cbfa1 and OSX in the Ins2 Akita/+ ;Tie2-Gfp tg mice. Scale bars, 100 μm. DAPI (blue) was used to visualize nuclei. Non-specific IgG control showed no staining. Vessel lumen faces upwards in the photos.
    Cd31, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 93/100, based on 727 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Thermo Fisher cd31 pecam 1 monoclonal antibody
    Endothelium contributes cells to aortic calcification of Ins2 Akita/+ mice (A) Aortic expression of endothelial markers <t>CD31</t> and vWF in Ins2 Akita/+ mice visualized by immunostaining. (B) Immunostaining of aortic tissues from wild type (WT) and Ins2 Akita/+ mice showed co-expression of endothelial markers CD31 (left) and vWF (right) and osteogenic markers Cbfa1 and Osterix (OSX) in the Ins2 Akita/+ mice. (C) Co-expression of CD31 and Cbfa1 in enzymatically dispersed aortic cells from WT and Ins2 Akita/+ mice , as determined by FACS. (D) Aortic expression of GFP in Tie2-Gfp tg and Ins2 Akita/+ ;Tie2-Gfp tg mice by immunostaining with anti-GFP antibodies. (E) Immunostaining of aortic tissues from Tie2-Gfp tg and Ins2 Akita/+ ;Tie2-Gfp tg mice showed co-expression of GFP with Cbfa1 and OSX in the Ins2 Akita/+ ;Tie2-Gfp tg mice. Scale bars, 100 μm. DAPI (blue) was used to visualize nuclei. Non-specific IgG control showed no staining. Vessel lumen faces upwards in the photos.
    Cd31 Pecam 1 Monoclonal Antibody, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 91/100, based on 800 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Abcam rabbit anti cd31
    Histological analysis of tissue from the pig esophagus at 2.5 months post implantation of a cellularized scaffold. ( A ) Macroscopic image of excised esophagus (proximal left, distal right). Samples of tissue were excised to include the site of surgery, monitored by endoscopy, with adjacent distal and proximal tissues for histology (box). ( B – F ) Representative images of hematoxylin and eosin ( B – E ) and Masson’s trichrome ( F – H ) stained excised esophageal tissue sections: ( C , F ) are at the center of the specimen, ( D , G ) are in the distal region at the anastomosis and ( E , H ) are from the same region of an esophagus without surgery (normal control). ( I – T ). Immunoreactivity for aSMA ( I , J ); Ki67 ( K , L ) suggests continued proliferation of mucosal and submucosal cells; presence of smooth muscle myosin heavy chain (SM-MHC) ( M , N ); <t>CD31</t> ( O , P ); transgelin/SM22a ( Q , R ); and the relative absence of striated myosin heavy chain ( S , T ) in tissue at the site of surgery. Scale bars: A = 6 cm, C-H = 200 μm, I-T = 200 μm.
    Rabbit Anti Cd31, supplied by Abcam, used in various techniques. Bioz Stars score: 99/100, based on 889 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Agilent technologies anti cd31
    Vascularized TOC devices permit robust growth of colorectal and pancreatic cancer cells. Three colorectal cancer (CRC268, CRC663, and CRC1180) and two pancreatic ductal adenocarcinoma cell lines (PDAC162 and PDAC175) were introduced into the side chambers with pre-formed vasculature in the central chamber. A: Representative immune fluorescence images taken at Days 2 and 9 after tumor cells were introduced. Green: GFP labeled endothelial cells formed vasculature. Red: RFP labelled tumor cells. The white dotted lines indicate the top and bottom boundaries of the central chamber. B: Quantification of angiogenesis, tumor cell proliferation, and migration/invasion on Day 9. Eight devices per tumor cell line were used for quantification. C: IHC images on parental CRC and PDAC tumor sections stained with <t>anti-CD31.</t> Representative images are shown at 100x and 200x magnification. D: Quantification of CD31-positive areas in parental tumors using IHC images. * p
    Anti Cd31, supplied by Agilent technologies, used in various techniques. Bioz Stars score: 93/100, based on 754 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Becton Dickinson cd31 fitc
    Surface markers of synovial MSCs before and 48 h after preservation. a Representative histogram of synovial MSCs by CD44-PE-Cy7, CD73-V450, CD90-PE, CD105-APC, and CD34-PE-Cy5. b Representative dot plot of synovial MSCs by <t>CD31-FITC</t> and CD45-APC-H7 as negative markers. c Calculated live cell number per tube (2 million synovial MSCs). Median values and interquartile ranges are shown ( n = 4). ND not detected
    Cd31 Fitc, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 93/100, based on 956 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Santa Cruz Biotechnology anti cd31
    Characterization of the P6C cells. (A) Holoclone formation of the P6C cells. Cells from primary colorectal cancer tissue and paired normal colon tissue were trypsinized and seeded into a 6-well plate. A primary clone from a single cancer cell and a holoclone of the P6C cells (passage 20) are shown in the top column, respectively. A representative primary sphere from the colon cancer tissue and dispersed normal colon cells are shown in the bottom column. Scale bars, 200 μm. (B) Expression of distinct markers in the P6C cells. Suspended P6C cells were incubated with anti-CD45-FITC, <t>CD31-FITC,</t> and CD24-FITC, respectively, and were analysed by flow cytometry. Donkey anti-mouse-FITC was used as a control. M1, negative; M2, positive. (C) Surface expression of CD44 in P6C cells as detected by immunofluorescence. P6C cells were grown attached to plates for 5 d, fixed with paraformaldehyde and incubated with anti-CD44 antibody. DAPI was used to stain the nucleus. Scale bar, 100 μm. (D) Relative expression levels of distinct markers under different culture conditions, including a P6C sphere, a P6C clone and SW480 cells. Suspended cells were collected and incubated with FITC conjugated antibodies and were analysed by flow cytometry. Each sample was analysed in triplicate, and the experiment was repeated 3 times. b P
    Anti Cd31, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 93/100, based on 866 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Dianova cd31
    Histopathology analysis of tumors reveals antibody-labeled nanoparticles localize to regions rich with immune cells. ( A ) Analysis of Prussian blue–positive (nanoparticle-rich) areas of tumors from nude mice injected with BH nanoparticles reveals only weak correlation with HER2 expression. ( B ) Conversely, this correlation is stronger in tumors from NSG mice. ( C and D ) Weak or no correlation was observed between BH nanoparticle presence and <t>CD31</t> + (vascular endothelium) regions. ( E ) Representative histology images of sequential sections showing IBA-1 + cells associated with Prussian blue–positive areas in HCC1954 (HER2 + ) tumors grown in NSG mice and treated with BH (a) hematoxylin and eosin (H E), (b) Prussian blue, (c) HER2 IHC, (d) IBA-1 IHC, (e) CD-31 IHC, (f) H E of another area from same tumor, (g) sequential section stained for Prussian blue shows positive staining for iron nanoparticles, and (h) immunofluorescence (IF) staining for IBA-1 shows positivity in the nanoparticle accumulated region. ( F and G ) Iron recovery from HER2 + (HCC1954) or HER2 − (MDA-MB-231) tumors is similar whether BNF nanoparticles have trastuzumab (anti-HER2) or human IgG (polyclonal), suggesting that antibody-antigen binding does not drive intratumor nanoparticle accumulation. ns, not statistically significant.
    Cd31, supplied by Dianova, used in various techniques. Bioz Stars score: 94/100, based on 604 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    R&D Systems mouse rat cd31 pecam 1 antibody
    Histopathology analysis of tumors reveals antibody-labeled nanoparticles localize to regions rich with immune cells. ( A ) Analysis of Prussian blue–positive (nanoparticle-rich) areas of tumors from nude mice injected with BH nanoparticles reveals only weak correlation with HER2 expression. ( B ) Conversely, this correlation is stronger in tumors from NSG mice. ( C and D ) Weak or no correlation was observed between BH nanoparticle presence and <t>CD31</t> + (vascular endothelium) regions. ( E ) Representative histology images of sequential sections showing IBA-1 + cells associated with Prussian blue–positive areas in HCC1954 (HER2 + ) tumors grown in NSG mice and treated with BH (a) hematoxylin and eosin (H E), (b) Prussian blue, (c) HER2 IHC, (d) IBA-1 IHC, (e) CD-31 IHC, (f) H E of another area from same tumor, (g) sequential section stained for Prussian blue shows positive staining for iron nanoparticles, and (h) immunofluorescence (IF) staining for IBA-1 shows positivity in the nanoparticle accumulated region. ( F and G ) Iron recovery from HER2 + (HCC1954) or HER2 − (MDA-MB-231) tumors is similar whether BNF nanoparticles have trastuzumab (anti-HER2) or human IgG (polyclonal), suggesting that antibody-antigen binding does not drive intratumor nanoparticle accumulation. ns, not statistically significant.
    Mouse Rat Cd31 Pecam 1 Antibody, supplied by R&D Systems, used in various techniques. Bioz Stars score: 99/100, based on 554 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Bio-Rad cd31
    Myocardial histology. ( A ) Representative image of histology sampling from infarcted areas ( B , C , E ); peri-infarcted areas ( D , F , G ); ( B ) Masson’s Trichrome ×1.25 ( C ) Hematoxylin-eosin ×5 ( D ) Integrin α v β 3 ×20 ( E ) α-smooth muscle actin ×20 ( F ) CD68 ×20 ( G ) <t>CD31</t> ×20 ( H ) Percentage fibrosis in all sample areas (I) α-smooth muscle actin percentage in scar ( J ) CD31 vessel density per high power field. pASC: porcine adipose-derived stromal cells in alginate hydrogel, Arrows show co-localization of integrin α v β 3 and CD31.
    Cd31, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 92/100, based on 709 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Pharmingen cd31
    Keratin, Ezh2 and <t>CD31</t> expression in tumors treated with SKI-606 for 14 days. Luminal epithelial lineages (K8, red) and basal-like epithelia (K14, green) were visualized in sections of tumors of mice treated with vehicle (A-a) or SKI606. (A-b). B, localization of Ezh2 and E-cadherin in tumors. Lower panels show decreased Ezh2 expression of SKI-606-treated tumors. C, CD31 (red) staining of tumor sections. SKI-606-treated tumor (right) has more organized and regular spacing. D, image analysis reveals length of vessel perimeter (left), endothelium area of vessel (middle), by t-test and vessel density (right). *P
    Cd31, supplied by Pharmingen, used in various techniques. Bioz Stars score: 93/100, based on 578 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Representative figures of cellular atypia (A), CD31 expression levels (B) and vWF expression levels (C). Inserted images are the magnified views of each figures. Rectangle images at the bottom of (B) and (C) are immunohistochemical reactivity of CD31 and vWF in normal endothelial cells in the same slides of above images. Bars=20 µ m.

    Journal: The Journal of Veterinary Medical Science

    Article Title: Cellular atypia is negatively correlated with immunohistochemical reactivity of CD31 and vWF expression levels in canine hemangiosarcoma

    doi: 10.1292/jvms.17-0561

    Figure Lengend Snippet: Representative figures of cellular atypia (A), CD31 expression levels (B) and vWF expression levels (C). Inserted images are the magnified views of each figures. Rectangle images at the bottom of (B) and (C) are immunohistochemical reactivity of CD31 and vWF in normal endothelial cells in the same slides of above images. Bars=20 µ m.

    Article Snippet: Sections were incubated overnight at 4°C in anti-CD31 (anti-human CD31 mouse monoclonal antibody, clone JC/70A, 1:500; Thermo Fisher Scientific, Waltham, MA, U.S.A.) or anti-vWF (anti-human vWF rabbit polyclonal antibody, 1:1,000, Agilent technologies, Santa Clara, CA, U.S.A.) primary antibodies followed by secondary antibody reaction for 30 min at RT using biotinylated anti-mouse IgG + IgA + IgM (Nichirei biosciences) or biotinylated anti-rabbit IgG (Nichirei biosciences).

    Techniques: Expressing, Immunohistochemistry

    Association between (A) cellular atypia and CD31, (B) cellular atypia and vWF, and (C) CD31 and vWF. R: Pearson’s correlation coefficient. The immunohistochemical reactivity of CD31 and vWF were scored as negative=0, mild=1, moderate=2 and high=3. Cellular atypia levels were scored as mild=1, moderate=2 and severe=3.

    Journal: The Journal of Veterinary Medical Science

    Article Title: Cellular atypia is negatively correlated with immunohistochemical reactivity of CD31 and vWF expression levels in canine hemangiosarcoma

    doi: 10.1292/jvms.17-0561

    Figure Lengend Snippet: Association between (A) cellular atypia and CD31, (B) cellular atypia and vWF, and (C) CD31 and vWF. R: Pearson’s correlation coefficient. The immunohistochemical reactivity of CD31 and vWF were scored as negative=0, mild=1, moderate=2 and high=3. Cellular atypia levels were scored as mild=1, moderate=2 and severe=3.

    Article Snippet: Sections were incubated overnight at 4°C in anti-CD31 (anti-human CD31 mouse monoclonal antibody, clone JC/70A, 1:500; Thermo Fisher Scientific, Waltham, MA, U.S.A.) or anti-vWF (anti-human vWF rabbit polyclonal antibody, 1:1,000, Agilent technologies, Santa Clara, CA, U.S.A.) primary antibodies followed by secondary antibody reaction for 30 min at RT using biotinylated anti-mouse IgG + IgA + IgM (Nichirei biosciences) or biotinylated anti-rabbit IgG (Nichirei biosciences).

    Techniques: Immunohistochemistry

    Expression levels of CD31 (A) and vWF (B), and cellular atypia scores (C) in each growth pattern. Kruskal wallis test was done for statistical analysis. The immunohistochemical reactivity of CD31 and vWF were scored as negative=0, mild=1, moderate=2 and high=3. Cellular atypia levels were scored as mild=1, moderate=2 and severe=3.

    Journal: The Journal of Veterinary Medical Science

    Article Title: Cellular atypia is negatively correlated with immunohistochemical reactivity of CD31 and vWF expression levels in canine hemangiosarcoma

    doi: 10.1292/jvms.17-0561

    Figure Lengend Snippet: Expression levels of CD31 (A) and vWF (B), and cellular atypia scores (C) in each growth pattern. Kruskal wallis test was done for statistical analysis. The immunohistochemical reactivity of CD31 and vWF were scored as negative=0, mild=1, moderate=2 and high=3. Cellular atypia levels were scored as mild=1, moderate=2 and severe=3.

    Article Snippet: Sections were incubated overnight at 4°C in anti-CD31 (anti-human CD31 mouse monoclonal antibody, clone JC/70A, 1:500; Thermo Fisher Scientific, Waltham, MA, U.S.A.) or anti-vWF (anti-human vWF rabbit polyclonal antibody, 1:1,000, Agilent technologies, Santa Clara, CA, U.S.A.) primary antibodies followed by secondary antibody reaction for 30 min at RT using biotinylated anti-mouse IgG + IgA + IgM (Nichirei biosciences) or biotinylated anti-rabbit IgG (Nichirei biosciences).

    Techniques: Expressing, Immunohistochemistry

    Endothelial phenotype changes with stromal cell co-culture. a Endothelial staining of HS27a and b HS5 co-cultured vessels shows differences in (i) CD31, (ii) VE-cadherin (VE-Cad), and (iii) von Willebrand Factor (vWF). c RNA expression within the co-culture shows differences in endothelial junctional expression and inflammatory marker expression. * p

    Journal: Stem Cell Research & Therapy

    Article Title: Engineering a multicellular vascular niche to model hematopoietic cell trafficking

    doi: 10.1186/s13287-018-0808-2

    Figure Lengend Snippet: Endothelial phenotype changes with stromal cell co-culture. a Endothelial staining of HS27a and b HS5 co-cultured vessels shows differences in (i) CD31, (ii) VE-cadherin (VE-Cad), and (iii) von Willebrand Factor (vWF). c RNA expression within the co-culture shows differences in endothelial junctional expression and inflammatory marker expression. * p

    Article Snippet: RT-PCR on microvessels comparing the HS27a and HS5 stromal co-cultures to vessels with ECs alone showed a significant reduction in vWF (75% and 57% reduction in HS5 and HS27a vessels compared to EC-only vessels, respectively) and CD31 (59% and 58% reduction in expression in HS5 and HS27a vessels, respectively) expression in microvessels after co-culture.

    Techniques: Co-Culture Assay, Staining, Cell Culture, RNA Expression, Expressing, Marker

    Relationship between HSC-primed HECs and T1 pre-HSCs. (a) Representative FACS plots for sorting of the T1 pre-HSCs (CD31 + CD45 - CD41 low Kit + CD201 high ) from E11.0 AGM region of mouse embryos. Red box indicates the sampling cells for scRNA-seq. (b) Violin plots showing the expression levels of indicated genes in tif-HEC (including cluster HEC and PK44), T1 pre-HSC and lateAEC. (c) t-SNE plot of the cells included in the filtered initial dataset, PK44 dataset and T1 pre-HSC dataset, with clusters mapped on it. Cluster HEC and PK44 are combined as tif-HEC. (d) PCA plot of tif-HEC and T1 pre-HSC populations. (e) Enriched terms of PC2 positive and negative genes are shown, corresponding to the properties distinguishing tif-HEC and T1 pre-HSC, respectively. (f) Heatmap showing top 20 positive and negative genes of PC2. Genes were ordered by their contributions to PC2. (g) Trajectory of AEC clusters, tif-HEC and T1 pre-HSC inferred by Mpath. Arrows indicate the development directions predicted by sampling stages. (h) Representative FACS plots for sorting of the PK44 cells from E10.0 AGM region (left) and analysis of the immunophenotypic T1 pre-HSCs (right) after cultured in vitro for 4 days. (i) Representative CD31 immunostaining on the cultures of single T1 pre-HSCs from E11.0 AGM region, showing typical morphologies regarding distinct differentiation capacities. Cell frequencies of each type are also shown. Data are from 7 independent experiments with totally 89 embryos used. Scale bars, 400 μm.

    Journal: bioRxiv

    Article Title: Dissecting endothelial to haematopoietic stem cell transition by single-cell transcriptomic and functional analyses

    doi: 10.1101/2020.01.18.910356

    Figure Lengend Snippet: Relationship between HSC-primed HECs and T1 pre-HSCs. (a) Representative FACS plots for sorting of the T1 pre-HSCs (CD31 + CD45 - CD41 low Kit + CD201 high ) from E11.0 AGM region of mouse embryos. Red box indicates the sampling cells for scRNA-seq. (b) Violin plots showing the expression levels of indicated genes in tif-HEC (including cluster HEC and PK44), T1 pre-HSC and lateAEC. (c) t-SNE plot of the cells included in the filtered initial dataset, PK44 dataset and T1 pre-HSC dataset, with clusters mapped on it. Cluster HEC and PK44 are combined as tif-HEC. (d) PCA plot of tif-HEC and T1 pre-HSC populations. (e) Enriched terms of PC2 positive and negative genes are shown, corresponding to the properties distinguishing tif-HEC and T1 pre-HSC, respectively. (f) Heatmap showing top 20 positive and negative genes of PC2. Genes were ordered by their contributions to PC2. (g) Trajectory of AEC clusters, tif-HEC and T1 pre-HSC inferred by Mpath. Arrows indicate the development directions predicted by sampling stages. (h) Representative FACS plots for sorting of the PK44 cells from E10.0 AGM region (left) and analysis of the immunophenotypic T1 pre-HSCs (right) after cultured in vitro for 4 days. (i) Representative CD31 immunostaining on the cultures of single T1 pre-HSCs from E11.0 AGM region, showing typical morphologies regarding distinct differentiation capacities. Cell frequencies of each type are also shown. Data are from 7 independent experiments with totally 89 embryos used. Scale bars, 400 μm.

    Article Snippet: Cells were stained by the following antibodies: B220 (eBioscience, RA3-6B2), CD3 (eBioscience, 145-2C11), CD4 (eBioscience, GK1.5), CD8a (eBioscience, 53-6.7), CD31 (BD or BioLegend, MEC13.3), CD41 (BD or eBioscience, MWReg30), CD43 (BD, S7), CD44 (eBioscience or BioLegend, IM7), CD45.1 (eBioscience, A20), CD45.2 (eBioscience, 104), CD45 (eBioscience, 30-F11), CD144 (eBioscience, eBioBV13), CD201 (eBioscience, eBio1560), Flk1 (eBioscience, Avas12a1), Kit (eBioscience, 2B8), Ly-6G (BioLegend, 1A8), and Mac-1 (eBioscience, M1/70).

    Techniques: FACS, Sampling, Expressing, Cell Culture, In Vitro, Immunostaining

    Identifying Neurl3 as a signature gene of HSC-primed HECs validated by functional and transcriptomic evaluation. (a) Dot plot showing the average and percentage expression of HEC signature genes in the indicated clusters. Genes are ordered by their median expression level in tif-HEC. Pre-HSC si gnature genes are marked as aquamarine. (b) Schematic model of the gene targeting strategy for generating Neurl3 EGFP/+ reporter mouse line via CRISPR/Cas9 system. (c) Representative FACS analysis of the E10.0 AGM region in Neurl3 EGFP/+ embryos, FACS plot to the right showing PK44 cells (red dots) mapped on it. (d) Representative FACS plot for sorting of the indicated cell populations from E10.0 caudal half of Neurl3 EGFP/+ embryos. (e) Graph showing the donor chimerism at 16 weeks after transplantation of the derivatives of the indicated populations from the caudal half of E10.0 Neurl3 EGFP/+ embryos. (f) Graph showing the donor chimerism at 4-16 weeks post-transplantation. The recipients were transplanted with the derivatives of CD41 - CD43 - CD45 - CD31 + CD44 + Neurl3-EGFP + population from the caudal half of E10.0 Neurl3 EGFP/+ embryos. Number of repopulated/total recipients is shown in the brackets. (g) t-SNE plot of the cells included in the filtered initial dataset and additional PK44 and NE+ datasets, with clusters mapped on it. Cluster HEC and PK44 are combined as tif-HEC. NE+, CD41 - CD43 - CD45 - CD31 + CD44 + Neurl3-EGFP + population from E10.0 AGM region. (h) Dot plot showing the average and percentage expression of selected HEC feature genes in the indicated clusters. Pre-HSC signature genes are marked as aquamarine. (i) Heatmap showing the correlation coefficient between each two clusters with hierarchical clustering using average method. Pearson correlation coefficient is calculated using average expression of highly variable genes in each cluster.

    Journal: bioRxiv

    Article Title: Dissecting endothelial to haematopoietic stem cell transition by single-cell transcriptomic and functional analyses

    doi: 10.1101/2020.01.18.910356

    Figure Lengend Snippet: Identifying Neurl3 as a signature gene of HSC-primed HECs validated by functional and transcriptomic evaluation. (a) Dot plot showing the average and percentage expression of HEC signature genes in the indicated clusters. Genes are ordered by their median expression level in tif-HEC. Pre-HSC si gnature genes are marked as aquamarine. (b) Schematic model of the gene targeting strategy for generating Neurl3 EGFP/+ reporter mouse line via CRISPR/Cas9 system. (c) Representative FACS analysis of the E10.0 AGM region in Neurl3 EGFP/+ embryos, FACS plot to the right showing PK44 cells (red dots) mapped on it. (d) Representative FACS plot for sorting of the indicated cell populations from E10.0 caudal half of Neurl3 EGFP/+ embryos. (e) Graph showing the donor chimerism at 16 weeks after transplantation of the derivatives of the indicated populations from the caudal half of E10.0 Neurl3 EGFP/+ embryos. (f) Graph showing the donor chimerism at 4-16 weeks post-transplantation. The recipients were transplanted with the derivatives of CD41 - CD43 - CD45 - CD31 + CD44 + Neurl3-EGFP + population from the caudal half of E10.0 Neurl3 EGFP/+ embryos. Number of repopulated/total recipients is shown in the brackets. (g) t-SNE plot of the cells included in the filtered initial dataset and additional PK44 and NE+ datasets, with clusters mapped on it. Cluster HEC and PK44 are combined as tif-HEC. NE+, CD41 - CD43 - CD45 - CD31 + CD44 + Neurl3-EGFP + population from E10.0 AGM region. (h) Dot plot showing the average and percentage expression of selected HEC feature genes in the indicated clusters. Pre-HSC signature genes are marked as aquamarine. (i) Heatmap showing the correlation coefficient between each two clusters with hierarchical clustering using average method. Pearson correlation coefficient is calculated using average expression of highly variable genes in each cluster.

    Article Snippet: Cells were stained by the following antibodies: B220 (eBioscience, RA3-6B2), CD3 (eBioscience, 145-2C11), CD4 (eBioscience, GK1.5), CD8a (eBioscience, 53-6.7), CD31 (BD or BioLegend, MEC13.3), CD41 (BD or eBioscience, MWReg30), CD43 (BD, S7), CD44 (eBioscience or BioLegend, IM7), CD45.1 (eBioscience, A20), CD45.2 (eBioscience, 104), CD45 (eBioscience, 30-F11), CD144 (eBioscience, eBioBV13), CD201 (eBioscience, eBio1560), Flk1 (eBioscience, Avas12a1), Kit (eBioscience, 2B8), Ly-6G (BioLegend, 1A8), and Mac-1 (eBioscience, M1/70).

    Techniques: Functional Assay, Expressing, CRISPR, FACS, Transplantation Assay

    Identification of the HSC-competent and endothelial-haematopoietic bi-potent HECs. (a) Detailed information of the co-culture/transplantation assays performed with E9.5-E10.0 caudal half cells. (b) Blood chimerism of the primary and secondary recipients at 4-16 weeks post-transplantation. The primary recipients were transplanted with the derivatives of the indicated cell populations from the caudal half of E9.5-E10.0 embryos. The paired primary and corresponding secondary repopulated mice are show as the same symbol and color. Numbers of repopulated/total recipients are shown in the brackets. Only the recipients survived to 16 weeks post-transplantation are shown. (c) FACS plots showing representative primary recipients with long-term (16 weeks), multi-organ and multi-lineage repopulations transplanted with the derivatives of the indicated cell populations from the caudal half of E9.5-E10.0 embryos. Donor-derived (CD45.1+CD45.2+) myeloid (Gr-1+/Mac-1+), B lymphoid (B220+), and T lymphoid (CD3+) cells in multiple haematopoietic organs are shown. (d) Heatmap showing the expression of selected genes in earlyAEC, lateAEC, HEC and PK44 populations. Note the similarity of expression patterns between HEC and PK44. (e) Graph showing the endothelial potential of different cell populations in E9.5-E10.0 body part of embryo proper. Cells with indicated immunophenotype were isolated by FACS, co-cultured with OP9 stromal cells for 7 days, and stained with CD31 to identify the endothelial tubes. Data are means ± s.d.. For E9.5 embryos, data are from 3 independent experiments with 6-9 embryo equivalents pooled for each experiment. For E10.0 embryos, data are from 3 independent experiments with 8-9 embryo equivalents pooled for each experiment. (f) Detailed information of endothelial-haematopoietic bi-potential induction assays performed with cells from E9.5-E10.0 caudal half or AGM region.

    Journal: bioRxiv

    Article Title: Dissecting endothelial to haematopoietic stem cell transition by single-cell transcriptomic and functional analyses

    doi: 10.1101/2020.01.18.910356

    Figure Lengend Snippet: Identification of the HSC-competent and endothelial-haematopoietic bi-potent HECs. (a) Detailed information of the co-culture/transplantation assays performed with E9.5-E10.0 caudal half cells. (b) Blood chimerism of the primary and secondary recipients at 4-16 weeks post-transplantation. The primary recipients were transplanted with the derivatives of the indicated cell populations from the caudal half of E9.5-E10.0 embryos. The paired primary and corresponding secondary repopulated mice are show as the same symbol and color. Numbers of repopulated/total recipients are shown in the brackets. Only the recipients survived to 16 weeks post-transplantation are shown. (c) FACS plots showing representative primary recipients with long-term (16 weeks), multi-organ and multi-lineage repopulations transplanted with the derivatives of the indicated cell populations from the caudal half of E9.5-E10.0 embryos. Donor-derived (CD45.1+CD45.2+) myeloid (Gr-1+/Mac-1+), B lymphoid (B220+), and T lymphoid (CD3+) cells in multiple haematopoietic organs are shown. (d) Heatmap showing the expression of selected genes in earlyAEC, lateAEC, HEC and PK44 populations. Note the similarity of expression patterns between HEC and PK44. (e) Graph showing the endothelial potential of different cell populations in E9.5-E10.0 body part of embryo proper. Cells with indicated immunophenotype were isolated by FACS, co-cultured with OP9 stromal cells for 7 days, and stained with CD31 to identify the endothelial tubes. Data are means ± s.d.. For E9.5 embryos, data are from 3 independent experiments with 6-9 embryo equivalents pooled for each experiment. For E10.0 embryos, data are from 3 independent experiments with 8-9 embryo equivalents pooled for each experiment. (f) Detailed information of endothelial-haematopoietic bi-potential induction assays performed with cells from E9.5-E10.0 caudal half or AGM region.

    Article Snippet: Cells were stained by the following antibodies: B220 (eBioscience, RA3-6B2), CD3 (eBioscience, 145-2C11), CD4 (eBioscience, GK1.5), CD8a (eBioscience, 53-6.7), CD31 (BD or BioLegend, MEC13.3), CD41 (BD or eBioscience, MWReg30), CD43 (BD, S7), CD44 (eBioscience or BioLegend, IM7), CD45.1 (eBioscience, A20), CD45.2 (eBioscience, 104), CD45 (eBioscience, 30-F11), CD144 (eBioscience, eBioBV13), CD201 (eBioscience, eBio1560), Flk1 (eBioscience, Avas12a1), Kit (eBioscience, 2B8), Ly-6G (BioLegend, 1A8), and Mac-1 (eBioscience, M1/70).

    Techniques: Co-Culture Assay, Transplantation Assay, Mouse Assay, FACS, Derivative Assay, Expressing, Isolation, Cell Culture, Staining

    In situ localization and in vitro function of the dynamic HECs marked by Neurl3-EGFP reporter. (a) Representative immunostaining on cross sections at AGM region of E9.5 (upper), E10.0 (middle) and E10.5 (lower) Neurl3 EGFP/+ embryos. Arrows indicate Neurl3 + aortic ECs. Yellow arrowheads indicate Neurl3 + bulging and bulged cells and IAHCs. Aquamarine arrowheads indicate CD44 + Runx1 + Neurl3 - haematopoietic cells distributed outside the aorta. nt, neural tube; DA, dorsal aorta. Scale bars, 100 μm. (b) Representative FACS analysis of the E10.0 AGM region of Neurl3 EGFP/+ embryos. FACS plots to the right showing PK44 cells (red dots, upper) and CD31 + Kit high cells (blue dots, lower) mapped on, respectively, with their contributions to each gated population indicated. (c) Representative CD31 and CD45 immunostaining on the cultures of single CD41 - CD43 - CD45 - CD31 + CD44 + Neurl3-EGFP + cells from E10.0 AGM region of Neurl3 EGFP/+ embryos, showing typical morphologies regarding distinct differentiation potentials. Cell frequencies of each kind of potential are also shown. Data are from 5 independent experiments with totally 37 embryos used. Scale bars, 400 μm. (d) Column charts showing the proportions of positive wells in the indicated populations (lower) for each kind of potential. The experiments were performed with CD41 - CD43 - CD45 - CD31 + CD44 - Neurl3-EGFP + single cells from E9.5 caudal half or E10.0-E10.5 AGM region of Neurl3 EGFP/+ embryos with PK44 indexed. Progenies from PK44 and non-PK44 cells are represented by distinct fill patterns. (e) Expression of CD44 and Neurl3-EGFP and values of FSC-A and SSC-A in the index-sorted single CD41 - CD43 - CD45 - CD31 + CD44 + Neurl3-EGFP + cells with differentiation potential based on in vitro functional evaluation. Cells with different kinds of potential are mapped onto the reference FACS plots (grey dots). Solid boxes (left of each stage) indicate the gates of the populations for FACS sorting. The enlarged views of solid boxes are shown below.

    Journal: bioRxiv

    Article Title: Dissecting endothelial to haematopoietic stem cell transition by single-cell transcriptomic and functional analyses

    doi: 10.1101/2020.01.18.910356

    Figure Lengend Snippet: In situ localization and in vitro function of the dynamic HECs marked by Neurl3-EGFP reporter. (a) Representative immunostaining on cross sections at AGM region of E9.5 (upper), E10.0 (middle) and E10.5 (lower) Neurl3 EGFP/+ embryos. Arrows indicate Neurl3 + aortic ECs. Yellow arrowheads indicate Neurl3 + bulging and bulged cells and IAHCs. Aquamarine arrowheads indicate CD44 + Runx1 + Neurl3 - haematopoietic cells distributed outside the aorta. nt, neural tube; DA, dorsal aorta. Scale bars, 100 μm. (b) Representative FACS analysis of the E10.0 AGM region of Neurl3 EGFP/+ embryos. FACS plots to the right showing PK44 cells (red dots, upper) and CD31 + Kit high cells (blue dots, lower) mapped on, respectively, with their contributions to each gated population indicated. (c) Representative CD31 and CD45 immunostaining on the cultures of single CD41 - CD43 - CD45 - CD31 + CD44 + Neurl3-EGFP + cells from E10.0 AGM region of Neurl3 EGFP/+ embryos, showing typical morphologies regarding distinct differentiation potentials. Cell frequencies of each kind of potential are also shown. Data are from 5 independent experiments with totally 37 embryos used. Scale bars, 400 μm. (d) Column charts showing the proportions of positive wells in the indicated populations (lower) for each kind of potential. The experiments were performed with CD41 - CD43 - CD45 - CD31 + CD44 - Neurl3-EGFP + single cells from E9.5 caudal half or E10.0-E10.5 AGM region of Neurl3 EGFP/+ embryos with PK44 indexed. Progenies from PK44 and non-PK44 cells are represented by distinct fill patterns. (e) Expression of CD44 and Neurl3-EGFP and values of FSC-A and SSC-A in the index-sorted single CD41 - CD43 - CD45 - CD31 + CD44 + Neurl3-EGFP + cells with differentiation potential based on in vitro functional evaluation. Cells with different kinds of potential are mapped onto the reference FACS plots (grey dots). Solid boxes (left of each stage) indicate the gates of the populations for FACS sorting. The enlarged views of solid boxes are shown below.

    Article Snippet: Cells were stained by the following antibodies: B220 (eBioscience, RA3-6B2), CD3 (eBioscience, 145-2C11), CD4 (eBioscience, GK1.5), CD8a (eBioscience, 53-6.7), CD31 (BD or BioLegend, MEC13.3), CD41 (BD or eBioscience, MWReg30), CD43 (BD, S7), CD44 (eBioscience or BioLegend, IM7), CD45.1 (eBioscience, A20), CD45.2 (eBioscience, 104), CD45 (eBioscience, 30-F11), CD144 (eBioscience, eBioBV13), CD201 (eBioscience, eBio1560), Flk1 (eBioscience, Avas12a1), Kit (eBioscience, 2B8), Ly-6G (BioLegend, 1A8), and Mac-1 (eBioscience, M1/70).

    Techniques: In Situ, In Vitro, Immunostaining, FACS, Expressing, Functional Assay

    DMOG treatment decreased apoptotic cells and increased neovascularization. a Representative co-staining of TUNEL-positive ( bright green nuclei marked by arrows ) and DAPI+ ( blue nuclei ) cells in non-infarcted and infarcted mice ± DMOG treatment 30 days after MI. Scale bar represents 25 μM. b Representative immunohistochemical staining of CD31 ( brown ) in infarcted hearts 30 days after MI. Scale bar represents 25 μM. c Bar graph representing the percentage of TUNEL-positive nuclei to total nuclei in the infarct border zone 30 days after MI. Data represent mean ± SD ( n = 6). d Bar graph showing the numbers of CD31+ capillaries in the infarct border zone in non-infarcted control mice, infarcted mice, and infarcted mice treated with DMOG 30 days after MI. Data represent mean ± SD ( n = 6); ** p

    Journal: Journal of Molecular Medicine (Berlin, Germany)

    Article Title: Prolyl-hydroxylase inhibition induces SDF-1 associated with increased CXCR4+/CD11b+ subpopulations and cardiac repair

    doi: 10.1007/s00109-017-1543-3

    Figure Lengend Snippet: DMOG treatment decreased apoptotic cells and increased neovascularization. a Representative co-staining of TUNEL-positive ( bright green nuclei marked by arrows ) and DAPI+ ( blue nuclei ) cells in non-infarcted and infarcted mice ± DMOG treatment 30 days after MI. Scale bar represents 25 μM. b Representative immunohistochemical staining of CD31 ( brown ) in infarcted hearts 30 days after MI. Scale bar represents 25 μM. c Bar graph representing the percentage of TUNEL-positive nuclei to total nuclei in the infarct border zone 30 days after MI. Data represent mean ± SD ( n = 6). d Bar graph showing the numbers of CD31+ capillaries in the infarct border zone in non-infarcted control mice, infarcted mice, and infarcted mice treated with DMOG 30 days after MI. Data represent mean ± SD ( n = 6); ** p

    Article Snippet: Cells were incubated for 40 min in the dark at 4 °C with the following fluoresceinisothiocyanate (FITC)-, phycoerythrin (PE)-, and peridininchlorophyll-protein (PerCP)-conjugated monoclonal antibodies: CD45-PerCP, CD11b-PerCP, CD11b-PE, CD4-PE, CD20-PE, CD31-PE, CD34-PE, Flk-PE, CD86-PE, CD206-PE, F4/80-PE, CD133-PE, c-kit-PE, Sca-1-PE, CD3-biotin, CD45R/B220-biotin, CD11b-biotin, TER-119-biotin, and Ly-6G-biotin (all from BD Pharmingen).

    Techniques: Staining, TUNEL Assay, Mouse Assay, Immunohistochemistry

    Increased tumor hypoxia in obese mice. Tumors in mice fed a high-fat diet (HFD) have higher Ki67+ counts ( a , scalebar 200 um). CD31 immunohistochemical analysis shows lower vessel density in tumors from HFD mice ( b , scalebar 200 uM). This is supported by fluorescence-activated cell sorting quantification ( c ). Pimonidazole (PIMO) staining in mouse fed a regular diet (RD) or a HFD demonstrate greater hypoxic areas in tumors from HFD mice ( d , scalebar 200 uM). Quantitative PCR analyses on RD and HFD tumors show upregulation of hypoxia inducible factor 1 alpha (HIF1α) targets ( e , n = 5) and faster tumor progression ( f , n = 5). Arbp, attachment region binding protein

    Journal: Breast Cancer Research : BCR

    Article Title: Obesity promotes the expansion of metastasis-initiating cells in breast cancer

    doi: 10.1186/s13058-018-1029-4

    Figure Lengend Snippet: Increased tumor hypoxia in obese mice. Tumors in mice fed a high-fat diet (HFD) have higher Ki67+ counts ( a , scalebar 200 um). CD31 immunohistochemical analysis shows lower vessel density in tumors from HFD mice ( b , scalebar 200 uM). This is supported by fluorescence-activated cell sorting quantification ( c ). Pimonidazole (PIMO) staining in mouse fed a regular diet (RD) or a HFD demonstrate greater hypoxic areas in tumors from HFD mice ( d , scalebar 200 uM). Quantitative PCR analyses on RD and HFD tumors show upregulation of hypoxia inducible factor 1 alpha (HIF1α) targets ( e , n = 5) and faster tumor progression ( f , n = 5). Arbp, attachment region binding protein

    Article Snippet: In agreement with this, we found fewer vessels and lower fractions of CD31+ cells in tumors in obese mice (Fig. and Additional file : Figure S2A).

    Techniques: Mouse Assay, Immunohistochemistry, Fluorescence, FACS, Staining, Real-time Polymerase Chain Reaction, Binding Assay

    Macroscopic and microscopic features of the plugs two and eight weeks after implantation. Plugs were harvested two ( A ) and eight weeks ( B ) after subcutaneous co-transplantation of ECFC+MSPC (ratio 80∶ 20, left column), MSPC only (2 nd column), ECFC only (3 rd column), and cell-free matrix (right column). Representative macroscopic pictures of plugs in subcutaneous location (upper rows), micrographs of hematoxylin and eosin staining (HE, middle rows) and immune histochemistry combining human CD31 colored brown with hematoxylin counter-stain (huCD31, bottom rows) are depicted. Total cell number within the plugs was 2×10 6 per 300 µL matrix (n = 3 per group and time course).

    Journal: PLoS ONE

    Article Title: Identification of an Effective Early Signaling Signature during Neo-Vasculogenesis In Vivo by Ex Vivo Proteomic Profiling

    doi: 10.1371/journal.pone.0066909

    Figure Lengend Snippet: Macroscopic and microscopic features of the plugs two and eight weeks after implantation. Plugs were harvested two ( A ) and eight weeks ( B ) after subcutaneous co-transplantation of ECFC+MSPC (ratio 80∶ 20, left column), MSPC only (2 nd column), ECFC only (3 rd column), and cell-free matrix (right column). Representative macroscopic pictures of plugs in subcutaneous location (upper rows), micrographs of hematoxylin and eosin staining (HE, middle rows) and immune histochemistry combining human CD31 colored brown with hematoxylin counter-stain (huCD31, bottom rows) are depicted. Total cell number within the plugs was 2×10 6 per 300 µL matrix (n = 3 per group and time course).

    Article Snippet: Slides were exposed to un-conjugated monoclonal mouse anti-human antibodies specifically binding vimentin (human mesodermal derived cell marker which can label mesenchymal and endothelial and hematopoietic cells, clone: V9, 0.78 µg/mL, Dako), CD31 (clone: JC70A, 5.15 µg/mL, Dako), and CD90 (clone: EPR3132, Abcam, Cambridge, MA, USA) for 30–60 min at room temperature (RT).

    Techniques: Transplantation Assay, Staining

    Loss of TSP-1 promotes tumor angiogenesis and platelet infiltration. (A-C) Tumors from WT and TSP-1 null mice were sectioned and stained with CD31 for microvessels, SMA for mature vessels, and CD42 for platelets. Quantification of staining per tumor is represented as mean±SEM ( n =4). * represents p

    Journal: bioRxiv

    Article Title: Platelet TSP-1 Controls Prostate Cancer-Induced Osteoclast Differentiation and Bone Marrow-Derived Cell Mobilization through TGFβ-1

    doi: 10.1101/2020.02.11.943860

    Figure Lengend Snippet: Loss of TSP-1 promotes tumor angiogenesis and platelet infiltration. (A-C) Tumors from WT and TSP-1 null mice were sectioned and stained with CD31 for microvessels, SMA for mature vessels, and CD42 for platelets. Quantification of staining per tumor is represented as mean±SEM ( n =4). * represents p

    Article Snippet: Sections were then fixed in 4% PFA and incubated with primary antibodies: anti-CD31 (1:50, BD Pharmigen; RRID: AB_393571), anti-CD42b (1:75, Millipore; RRID: AB_10712312) or anti-smooth muscle actin (SMA, 1:100, Abcam; RRID: AB_2223021).

    Techniques: Mouse Assay, Staining

    Platelet TGF-β1 is increased in TSP-1 null mice, and its deficiency inhibits progenitor cell mobilization and increases tumor angiogenesis. (A) Platelet TGF-β1 concentration was measured from control and RM1 tumor-bearing WT and TSP-1 null mice and represented as mean±SEM ( n =3). (B) RM1 tumors from TGF-β1 fl/fl and PF4-Cre + ;TGF-β1 fl/fl mice were weighed and represented as mean±SEM ( n =3-11). (C) Whole blood was collected and analyzed by flow cytometry for circulating progenitor cells using the markers CXCR4 and CD117. Representative quantification of positive cell numbers as mean percent of control ( n =2). (D) Tumors were sectioned and stained with CD31 for microvessels, SMA for mature vessels, and CD42 for platelets. Quantification of staining per tumor is represented as mean±SEM ( n =4). * represents p

    Journal: bioRxiv

    Article Title: Platelet TSP-1 Controls Prostate Cancer-Induced Osteoclast Differentiation and Bone Marrow-Derived Cell Mobilization through TGFβ-1

    doi: 10.1101/2020.02.11.943860

    Figure Lengend Snippet: Platelet TGF-β1 is increased in TSP-1 null mice, and its deficiency inhibits progenitor cell mobilization and increases tumor angiogenesis. (A) Platelet TGF-β1 concentration was measured from control and RM1 tumor-bearing WT and TSP-1 null mice and represented as mean±SEM ( n =3). (B) RM1 tumors from TGF-β1 fl/fl and PF4-Cre + ;TGF-β1 fl/fl mice were weighed and represented as mean±SEM ( n =3-11). (C) Whole blood was collected and analyzed by flow cytometry for circulating progenitor cells using the markers CXCR4 and CD117. Representative quantification of positive cell numbers as mean percent of control ( n =2). (D) Tumors were sectioned and stained with CD31 for microvessels, SMA for mature vessels, and CD42 for platelets. Quantification of staining per tumor is represented as mean±SEM ( n =4). * represents p

    Article Snippet: Sections were then fixed in 4% PFA and incubated with primary antibodies: anti-CD31 (1:50, BD Pharmigen; RRID: AB_393571), anti-CD42b (1:75, Millipore; RRID: AB_10712312) or anti-smooth muscle actin (SMA, 1:100, Abcam; RRID: AB_2223021).

    Techniques: Mouse Assay, Concentration Assay, Flow Cytometry, Staining

    IPP51 induces cell proliferation inhibition, mitotic arrest, apoptosis and angiogenesis inhibition in vivo A. - J. Representative images of sections from tumor xenografts obtained from control mice (left panel) or IPP51 treated mice (right panel). A. and B. correspond to a H E staining. Arrowheads show pyknotic (densely stained) and fragmented nuclei, indicating cell death. C. and D. showed result of TUNEL labeling (red) to measure apoptosis. E. and F. correspond to staining of Ki-67 (green), a proliferation marker. G. and H. are representative pictures of phosphorylated histone H3 staining (red), a mitotic marker. DNA in C. - H. sections was counterstained with Hoechst (blue). I and J were obtained after immunohistochemical labeling of CD31. K. - M. Tumor slides from control and IPP51 treated mice were visualized under microscope and TUNEL, Ki67 and phosphorylated histone H3 positive cells were quantified. Two slides per sample were analyzed, counting 500 cells per slide in randomly selected fields. Data represent the arithmetic mean ± SEM of 3 mice.

    Journal: Oncotarget

    Article Title: IPP51, a chalcone acting as a microtubule inhibitor with in vivo antitumor activity against bladder carcinoma

    doi:

    Figure Lengend Snippet: IPP51 induces cell proliferation inhibition, mitotic arrest, apoptosis and angiogenesis inhibition in vivo A. - J. Representative images of sections from tumor xenografts obtained from control mice (left panel) or IPP51 treated mice (right panel). A. and B. correspond to a H E staining. Arrowheads show pyknotic (densely stained) and fragmented nuclei, indicating cell death. C. and D. showed result of TUNEL labeling (red) to measure apoptosis. E. and F. correspond to staining of Ki-67 (green), a proliferation marker. G. and H. are representative pictures of phosphorylated histone H3 staining (red), a mitotic marker. DNA in C. - H. sections was counterstained with Hoechst (blue). I and J were obtained after immunohistochemical labeling of CD31. K. - M. Tumor slides from control and IPP51 treated mice were visualized under microscope and TUNEL, Ki67 and phosphorylated histone H3 positive cells were quantified. Two slides per sample were analyzed, counting 500 cells per slide in randomly selected fields. Data represent the arithmetic mean ± SEM of 3 mice.

    Article Snippet: Anti-CD31 antibody (clone MEC 13.3) was obtained from BD Pharmingen (Le Pont de Claix, France).

    Techniques: Inhibition, In Vivo, Mouse Assay, Staining, TUNEL Assay, Labeling, Marker, Immunohistochemistry, Microscopy

    RGC-32 inhibited angiogenesis and blood flow recovery after ischemia in vivo (A) RGC-32 protein expression increased after femoral artery ligation. (B) Infrared imaging from mice subjected to femoral artery ligation and retroviral injection of either pBMN or pBMN-RGC-32 on quadriceps. (C) Quantification of blood-flow recovery after treating either control or RGC-32 retrovirus. Data are means ± SEM n=8. (D) Sections of quadriceps from pBMN or RGC-32 o/e mice stained for CD31 (left) and quantification of capillaries (right).

    Journal: Circulation

    Article Title: Response Gene to Complement 32 (RGC-32), a novel hypoxia-regulated angiogenic inhibitor

    doi: 10.1161/CIRCULATIONAHA.108.841502

    Figure Lengend Snippet: RGC-32 inhibited angiogenesis and blood flow recovery after ischemia in vivo (A) RGC-32 protein expression increased after femoral artery ligation. (B) Infrared imaging from mice subjected to femoral artery ligation and retroviral injection of either pBMN or pBMN-RGC-32 on quadriceps. (C) Quantification of blood-flow recovery after treating either control or RGC-32 retrovirus. Data are means ± SEM n=8. (D) Sections of quadriceps from pBMN or RGC-32 o/e mice stained for CD31 (left) and quantification of capillaries (right).

    Article Snippet: Immunohistochemistry and immunofluorescence were performed with the CD31 antibody (BD Pharmingen) as described previously.

    Techniques: Flow Cytometry, In Vivo, Expressing, Ligation, Imaging, Mouse Assay, Injection, Staining

    Average cell populations for SVF constructs fixed after initial incubation and 48-hours of dynamic culture (n = 6). A paired t-test indicated no significant statistical difference between initial and 48-hour constructs for each of the CD90, CD31, and CD34 markers (P = 0.63, 0.68, and 0.46 respectively).

    Journal: The Journal of thoracic and cardiovascular surgery

    Article Title: An Exploratory Study into the Preparation and Evaluation of a “Same-Day” Adipose Stem Cell-Based TEVG

    doi: 10.1016/j.jtcvs.2018.05.120

    Figure Lengend Snippet: Average cell populations for SVF constructs fixed after initial incubation and 48-hours of dynamic culture (n = 6). A paired t-test indicated no significant statistical difference between initial and 48-hour constructs for each of the CD90, CD31, and CD34 markers (P = 0.63, 0.68, and 0.46 respectively).

    Article Snippet: Sections were incubated with 1:100 dilutions of either rabbit antihuman CD90 (ab133350, Abcam, Cambridge, UK), rabbit anti-human CD31 (ab28364, Abcam, Cambridge, UK), or rabbit anti-human CD34 (ab81289, Abcam, Cambridge, UK), to evaluate the presence of mesenchymal stem cell, endothelial cell, or endothelial progenitor cell populations respectively.

    Techniques: Construct, Incubation

    mTOR and MAPK pathways are activated in murine angiosarcoma ( A ) Western blots analyzing different endothelial cell markers in protein lysates prepared from HUVECs untreated (–) or treated (+) with VEGFA, total lung, endothelial cells isolated from lung using CD31 beads, tumors from both Trp53; Ptpn12 and Pten; Ptpn12 DKO mice, and tumors from TKO mice. ( B ) Western blots using the same protein lysates as in (A) but investigating the indicated signaling molecules. ( C ) Immunohistochemical staining on tissue sections from a representative tumor from each of the indicated mouse genotypes. Scale bar in the top right panel is 50 µm and applies to all pictures. ( D ) Anti-phosphotyrosine immunoprecipitations (IP) from control lung and TKO angiosarcoma protein lysates were then probed for PDGFR-β (left panel). The reciprocal immunoprecipitation and blotting experiment (middle panel) and total PDGFR-β comparisons (right panel) are shown. The input lane represents 20 µg of protein lysate from the tumor while IPs were performed from 500 µg protein lysate. ( E ) Tumors from the indicated mouse genotypes were stained by IHC for PDGFR-β. Scale bars are 50 µm.

    Journal: Oncotarget

    Article Title: Combined mTOR and MEK inhibition is an effective therapy in a novel mouse model for angiosarcoma

    doi: 10.18632/oncotarget.25345

    Figure Lengend Snippet: mTOR and MAPK pathways are activated in murine angiosarcoma ( A ) Western blots analyzing different endothelial cell markers in protein lysates prepared from HUVECs untreated (–) or treated (+) with VEGFA, total lung, endothelial cells isolated from lung using CD31 beads, tumors from both Trp53; Ptpn12 and Pten; Ptpn12 DKO mice, and tumors from TKO mice. ( B ) Western blots using the same protein lysates as in (A) but investigating the indicated signaling molecules. ( C ) Immunohistochemical staining on tissue sections from a representative tumor from each of the indicated mouse genotypes. Scale bar in the top right panel is 50 µm and applies to all pictures. ( D ) Anti-phosphotyrosine immunoprecipitations (IP) from control lung and TKO angiosarcoma protein lysates were then probed for PDGFR-β (left panel). The reciprocal immunoprecipitation and blotting experiment (middle panel) and total PDGFR-β comparisons (right panel) are shown. The input lane represents 20 µg of protein lysate from the tumor while IPs were performed from 500 µg protein lysate. ( E ) Tumors from the indicated mouse genotypes were stained by IHC for PDGFR-β. Scale bars are 50 µm.

    Article Snippet: Other antibodies were also used at 1:1000 except where indicated: anti-PTPN12 (Abcam, Cambridge, MA, USA; ab76492), anti-VEGFR1 (Abcam; ab32152), anti-VEGFR3 (Thermo Fisher Scientific; PA5-16871), anti-CD31 (Abcam; ab28364), anti-actin (Sigma-Aldrich; A5441, 1:10000), anti-VE-cadherin (Santa Cruz, Dallas, TX, USA; sc-6458), anti-pTyr 4G10 (Millipore, Billerica, MA, USA; 05-321), anti-pFAK (Sigma; F7926).

    Techniques: Western Blot, Isolation, Mouse Assay, Immunohistochemistry, Staining, Immunoprecipitation

    Angiogenic response in virus-induced scars. Endothelial CD31 expression (red) marks blood vessels, PDGFRα in green and lipid membranes in grey. (Left column, A, C, E, G, I, K) Overview, showing changes in vessel morphology and density in PDGF induced scars. (Right column, B, D, F, H, J, L) Asterisks mark big vessels with altered morphology. (M) Quantification of small vessels in the different experimental groups, normalized to the empty vector control. (N) Quantification of vessel size (area of cross-sectioned vessels) for the different experimental groups, normalized to the empty vector control. Analyses were performed on 3–4 mice per experimental group. E = empty vector; As = PDGF-A short ; Al = PDGF-A long ; B = PDGF-B; C = PDGF-C; D = PDGF-D. Scale bars are 100 μm.

    Journal: PLoS ONE

    Article Title: Isoform-Specific Modulation of Inflammation Induced by Adenoviral Mediated Delivery of Platelet-Derived Growth Factors in the Adult Mouse Heart

    doi: 10.1371/journal.pone.0160930

    Figure Lengend Snippet: Angiogenic response in virus-induced scars. Endothelial CD31 expression (red) marks blood vessels, PDGFRα in green and lipid membranes in grey. (Left column, A, C, E, G, I, K) Overview, showing changes in vessel morphology and density in PDGF induced scars. (Right column, B, D, F, H, J, L) Asterisks mark big vessels with altered morphology. (M) Quantification of small vessels in the different experimental groups, normalized to the empty vector control. (N) Quantification of vessel size (area of cross-sectioned vessels) for the different experimental groups, normalized to the empty vector control. Analyses were performed on 3–4 mice per experimental group. E = empty vector; As = PDGF-A short ; Al = PDGF-A long ; B = PDGF-B; C = PDGF-C; D = PDGF-D. Scale bars are 100 μm.

    Article Snippet: Primary antibodies used: rat-anti-mouse CD31, 1.25 μg/ml (553370, www.bdbiosciences.com ); Cy3-conjugated mouse-anti-human α-SMA, 2.8 μg/ml (C6198, www.sigmaaldrich.com ); anti-mouse PDGFRβ, 2.5 μg/ml (14-1402-82, www.ebioscience.com ); rabbit-anti NG2, 10 μg/ml (AB5320, www.millipore.com ); PINP, 7.93 μg/ml (Bode et al., Eur J Vasc Endovasc Surg 2002); goat-anti Nkx2.5, 10 μg/ml (AF2444, www.rndsystems.com ); rabbit-anti-mouse Ki-67, 3 μg/ml (ab15580, www.abcam.com ); rat-anti CD45, 0.125 μg/ml (610266, www.bdbiosciences.com ).

    Techniques: Expressing, Plasmid Preparation, Mouse Assay

    FGF-2-stimulated endothelial Pdgfb and Pdgfd expression in vivo and in vitro. a Pdgfb and Pdgfd mRNA levels in CD31 + ECs freshly isolated from T241-vector or T241–FGF-2 tumors ( n = 3 samples; n = 3 mice for each group). b Pdgfb and Pdgfd mRNA levels in CD31 + ECs freshly isolated from VT- or BGJ398-treated T241-vector and -FGF-2 tumors ( n = 3 samples; n = 3 mice for each group). c Pdgfb and Pdgfd mRNA levels in cultivated CD31 + ECs in response to FGF-2 stimulation ( n = 3 samples/group). d Pdgfb and Pdgfd mRNA levels in FGF-2-stimulated cultivated CD31 + ECs transfected with scrambled -RNA, siFgfr1 -RNA, or siFgfr2 -RNA ( n = 3 samples/group). All data as means ± S.E.M.; Student’s t test, * P

    Journal: Cell Discovery

    Article Title: Dual roles of endothelial FGF-2–FGFR1–PDGF-BB and perivascular FGF-2–FGFR2–PDGFRβ signaling pathways in tumor vascular remodeling

    doi: 10.1038/s41421-017-0002-1

    Figure Lengend Snippet: FGF-2-stimulated endothelial Pdgfb and Pdgfd expression in vivo and in vitro. a Pdgfb and Pdgfd mRNA levels in CD31 + ECs freshly isolated from T241-vector or T241–FGF-2 tumors ( n = 3 samples; n = 3 mice for each group). b Pdgfb and Pdgfd mRNA levels in CD31 + ECs freshly isolated from VT- or BGJ398-treated T241-vector and -FGF-2 tumors ( n = 3 samples; n = 3 mice for each group). c Pdgfb and Pdgfd mRNA levels in cultivated CD31 + ECs in response to FGF-2 stimulation ( n = 3 samples/group). d Pdgfb and Pdgfd mRNA levels in FGF-2-stimulated cultivated CD31 + ECs transfected with scrambled -RNA, siFgfr1 -RNA, or siFgfr2 -RNA ( n = 3 samples/group). All data as means ± S.E.M.; Student’s t test, * P

    Article Snippet: Single cells were stained with a rat anti-mouse CD31 antibody (553370; BD-Pharmingen) on ice for 45 min, followed by incubation for 30 min with a goat anti-rat Alex555 antibody (A21434; Invitrogen).

    Techniques: Expressing, In Vivo, In Vitro, Isolation, Plasmid Preparation, Mouse Assay, Transfection

    FGF-2-induced angiogenesis, pericyte recruitment, and tumor growth in vivo. a Tumor microvessel and pericyte contents. CD31 + endothelial cell (red) and NG2 + pericyte (green) signals in FGF-2 + and FGF-2 − tumors. Bar = 50 μm. b Quantification of microvessel density, vascular coverage by pericytes, and NG2 + pericyte area ( n = 7 random fields; n = 4 mice for each group). c CD31 + endothelial (red) and NG2 + pericyte (green) signals in matrigels containing FGF-2 - tumors with and without FGF-2 protein. Bar = 50 μm. d Quantification of vascular coverage by pericytes and NG2 + pericyte area in matrigels with and without FGF-2 protein ( n = 7 random fields; n = 4 mice for each group). e Tumor growth rates of scrambled- shRNA and Fgf2- shRNA-transfected FGF-2 + tumors ( n = 4–6 animals/group). f CD31 + endothelial (red) and NG2 + pericyte (green) contents in scrambled- shRNA and Fgf2- shRNA-transfected FGF-2 + tumors. Bar = 50 μm. g Quantification of microvascular density, vascular coverage by pericytes, and NG2 + pericyte area in scrambled -shRNA and Fgf2- shRNA-transfected FGF-2 + tumors ( n = 7 random fields; n = 4 mice for each group). h Tumor growth rates of T241-vector and T241-FGF-2 fibrosarcomas ( n = 6 animals/group). i CD31 + endothelial (red) and NG2 + pericyte (green) contents in T241-vector and -FGF-2 fibrosarcomas. Bar = 50 μm. j Quantification of microvessel density, vascular coverage by pericytes, and NG2 + pericyte area in T241-vector and T241–FGF-2 fibrosarcomas ( n = 7 random fields; n = 4 mice for each group). Vessels and pericytes were visualized using whole mount staining. All data as means ± S.E.M. * P

    Journal: Cell Discovery

    Article Title: Dual roles of endothelial FGF-2–FGFR1–PDGF-BB and perivascular FGF-2–FGFR2–PDGFRβ signaling pathways in tumor vascular remodeling

    doi: 10.1038/s41421-017-0002-1

    Figure Lengend Snippet: FGF-2-induced angiogenesis, pericyte recruitment, and tumor growth in vivo. a Tumor microvessel and pericyte contents. CD31 + endothelial cell (red) and NG2 + pericyte (green) signals in FGF-2 + and FGF-2 − tumors. Bar = 50 μm. b Quantification of microvessel density, vascular coverage by pericytes, and NG2 + pericyte area ( n = 7 random fields; n = 4 mice for each group). c CD31 + endothelial (red) and NG2 + pericyte (green) signals in matrigels containing FGF-2 - tumors with and without FGF-2 protein. Bar = 50 μm. d Quantification of vascular coverage by pericytes and NG2 + pericyte area in matrigels with and without FGF-2 protein ( n = 7 random fields; n = 4 mice for each group). e Tumor growth rates of scrambled- shRNA and Fgf2- shRNA-transfected FGF-2 + tumors ( n = 4–6 animals/group). f CD31 + endothelial (red) and NG2 + pericyte (green) contents in scrambled- shRNA and Fgf2- shRNA-transfected FGF-2 + tumors. Bar = 50 μm. g Quantification of microvascular density, vascular coverage by pericytes, and NG2 + pericyte area in scrambled -shRNA and Fgf2- shRNA-transfected FGF-2 + tumors ( n = 7 random fields; n = 4 mice for each group). h Tumor growth rates of T241-vector and T241-FGF-2 fibrosarcomas ( n = 6 animals/group). i CD31 + endothelial (red) and NG2 + pericyte (green) contents in T241-vector and -FGF-2 fibrosarcomas. Bar = 50 μm. j Quantification of microvessel density, vascular coverage by pericytes, and NG2 + pericyte area in T241-vector and T241–FGF-2 fibrosarcomas ( n = 7 random fields; n = 4 mice for each group). Vessels and pericytes were visualized using whole mount staining. All data as means ± S.E.M. * P

    Article Snippet: Single cells were stained with a rat anti-mouse CD31 antibody (553370; BD-Pharmingen) on ice for 45 min, followed by incubation for 30 min with a goat anti-rat Alex555 antibody (A21434; Invitrogen).

    Techniques: In Vivo, Mouse Assay, shRNA, Transfection, Plasmid Preparation, Staining

    Vascular perfusion and permeability in FGF-2 + and FGF-2 - tumors in vivo. a , c , e CD31 + tumor vasculature (red) and perfusion of 2000-kDa dextran (green) in FGF-2 + and FGF − , scrambled- shRNA and Fgf2- shRNA-transfected FGF-2 + , and T241-vector and T241-FGF-2 tumors. Yellow color indicates double positive signals and perfused vessels. Bar = 50 μm. b , d , f Quantification of blood perfusion in in FGF-2 + and FGF − , scrambled- shRNA and Fgf2- shRNA-transfected FGF-2 + , and T241-vector and T241-FGF-2 tumors ( n = 10 random fields; n = 3 mice for each group). g , i , k CD31 + tumor vasculature (red) and leakiness of 70-kDa dextran (green) in FGF-2 + and FGF − , scrambled- shRNA and Fgf2- shRNA-transfected FGF-2 + , and T241-vector and T241-FGF-2 tumors. Arrowheads indicate extravagated dextran (green). Intravascular dextran molecules are in yellow color. Bar = 50 μm. h , j , l Quantification of vascular permeability of 70-kDa dextran in FGF-2 + and FGF − , scrambled- shRNA and Fgf2- shRNA-transfected FGF-2 + , and T241-vector and T241-FGF-2 tumors ( n = 10 random fields; n = 3 mice for each group). Images are shown using whole mount staining. All data as means ± S.E.M.; Student’s t test, * P

    Journal: Cell Discovery

    Article Title: Dual roles of endothelial FGF-2–FGFR1–PDGF-BB and perivascular FGF-2–FGFR2–PDGFRβ signaling pathways in tumor vascular remodeling

    doi: 10.1038/s41421-017-0002-1

    Figure Lengend Snippet: Vascular perfusion and permeability in FGF-2 + and FGF-2 - tumors in vivo. a , c , e CD31 + tumor vasculature (red) and perfusion of 2000-kDa dextran (green) in FGF-2 + and FGF − , scrambled- shRNA and Fgf2- shRNA-transfected FGF-2 + , and T241-vector and T241-FGF-2 tumors. Yellow color indicates double positive signals and perfused vessels. Bar = 50 μm. b , d , f Quantification of blood perfusion in in FGF-2 + and FGF − , scrambled- shRNA and Fgf2- shRNA-transfected FGF-2 + , and T241-vector and T241-FGF-2 tumors ( n = 10 random fields; n = 3 mice for each group). g , i , k CD31 + tumor vasculature (red) and leakiness of 70-kDa dextran (green) in FGF-2 + and FGF − , scrambled- shRNA and Fgf2- shRNA-transfected FGF-2 + , and T241-vector and T241-FGF-2 tumors. Arrowheads indicate extravagated dextran (green). Intravascular dextran molecules are in yellow color. Bar = 50 μm. h , j , l Quantification of vascular permeability of 70-kDa dextran in FGF-2 + and FGF − , scrambled- shRNA and Fgf2- shRNA-transfected FGF-2 + , and T241-vector and T241-FGF-2 tumors ( n = 10 random fields; n = 3 mice for each group). Images are shown using whole mount staining. All data as means ± S.E.M.; Student’s t test, * P

    Article Snippet: Single cells were stained with a rat anti-mouse CD31 antibody (553370; BD-Pharmingen) on ice for 45 min, followed by incubation for 30 min with a goat anti-rat Alex555 antibody (A21434; Invitrogen).

    Techniques: Permeability, In Vivo, shRNA, Transfection, Plasmid Preparation, Mouse Assay, Staining

    FGFRs and PDGFRs in pericyte recruitment. a RT-PCR analysis of mRNA expression levels of FGF receptors in pericytes freshly isolated from T241-vector and T241–FGF-2 tumors using magnetic bead separation. Beta-actin serves as a control. b CD31 + endothelial (red) and NG2 + pericyte (green) signals in vehicle (VT)-, anti-FGFR1 neutralizing antibody-, anti-FGFR2 neutralizing antibody treated-, anti-FGFR3 neutralizing antibody-, BGJ398-, anti-PDGFRα neutralizing antibody-, anti-PDGFRβ neutralizing antibody-, and imatinib-treated FGF-2 + tumors. Arrowheads indicate pericyte-associated vessels. Images are presented using whole mount staining. Bar = 50 μm. c , d Quantification of NG2 + pericyte area versus the total CD31 + microvessels and vascular coverage. ( n = 7 random fields; n = 4 mice for each group). All data as means ± S.E.M.; Student’s t test, * P

    Journal: Cell Discovery

    Article Title: Dual roles of endothelial FGF-2–FGFR1–PDGF-BB and perivascular FGF-2–FGFR2–PDGFRβ signaling pathways in tumor vascular remodeling

    doi: 10.1038/s41421-017-0002-1

    Figure Lengend Snippet: FGFRs and PDGFRs in pericyte recruitment. a RT-PCR analysis of mRNA expression levels of FGF receptors in pericytes freshly isolated from T241-vector and T241–FGF-2 tumors using magnetic bead separation. Beta-actin serves as a control. b CD31 + endothelial (red) and NG2 + pericyte (green) signals in vehicle (VT)-, anti-FGFR1 neutralizing antibody-, anti-FGFR2 neutralizing antibody treated-, anti-FGFR3 neutralizing antibody-, BGJ398-, anti-PDGFRα neutralizing antibody-, anti-PDGFRβ neutralizing antibody-, and imatinib-treated FGF-2 + tumors. Arrowheads indicate pericyte-associated vessels. Images are presented using whole mount staining. Bar = 50 μm. c , d Quantification of NG2 + pericyte area versus the total CD31 + microvessels and vascular coverage. ( n = 7 random fields; n = 4 mice for each group). All data as means ± S.E.M.; Student’s t test, * P

    Article Snippet: Single cells were stained with a rat anti-mouse CD31 antibody (553370; BD-Pharmingen) on ice for 45 min, followed by incubation for 30 min with a goat anti-rat Alex555 antibody (A21434; Invitrogen).

    Techniques: Reverse Transcription Polymerase Chain Reaction, Expressing, Isolation, Plasmid Preparation, Staining, Mouse Assay

    Comparison of kappa values among pathologists for lymphovascular invasion (LVI) detection in colorectal cancers. While the average of LVI detection rate for each pathologist was 43% with hematoxylin and eosin (H E) only, 10% with CD31, 29% with D2-40, and 16% with ERG, the consensus reached 80% of LVI detection after a joint discussion about ERG patterns with LVI. a Interpreted by ERG

    Journal: Korean Journal of Pathology

    Article Title: ERG Immunohistochemistry as an Endothelial Marker for Assessing Lymphovascular Invasion

    doi: 10.4132/KoreanJPathol.2013.47.4.355

    Figure Lengend Snippet: Comparison of kappa values among pathologists for lymphovascular invasion (LVI) detection in colorectal cancers. While the average of LVI detection rate for each pathologist was 43% with hematoxylin and eosin (H E) only, 10% with CD31, 29% with D2-40, and 16% with ERG, the consensus reached 80% of LVI detection after a joint discussion about ERG patterns with LVI. a Interpreted by ERG

    Article Snippet: There have been reports of dual or triple immunohistochemical staining combining cytokeratin and vascular markers including CD31, CD34, and D2-40 being sensitive in detecting LVI than standardized H & E alone., It might be necessary to investigate dual or triple immunostaining including ERG to improve LVI detection.

    Techniques:

    Comparison of ERG, CD31, and D2-40 endothelial markers. (A) ERG, panendothelial marker showing nuclear immunoreactivity in artery, vein, and lymphatics. (B) ERG immunostaining specific for endothelial cells without cross-reactivity. (C) D31 immunostaining showing cross-reactivity in inflammatory cells. (D) D2-40 immunostaining showing cross-reactivity in fibroblasts.

    Journal: Korean Journal of Pathology

    Article Title: ERG Immunohistochemistry as an Endothelial Marker for Assessing Lymphovascular Invasion

    doi: 10.4132/KoreanJPathol.2013.47.4.355

    Figure Lengend Snippet: Comparison of ERG, CD31, and D2-40 endothelial markers. (A) ERG, panendothelial marker showing nuclear immunoreactivity in artery, vein, and lymphatics. (B) ERG immunostaining specific for endothelial cells without cross-reactivity. (C) D31 immunostaining showing cross-reactivity in inflammatory cells. (D) D2-40 immunostaining showing cross-reactivity in fibroblasts.

    Article Snippet: There have been reports of dual or triple immunohistochemical staining combining cytokeratin and vascular markers including CD31, CD34, and D2-40 being sensitive in detecting LVI than standardized H & E alone., It might be necessary to investigate dual or triple immunostaining including ERG to improve LVI detection.

    Techniques: Marker, Immunostaining

    Moesin is expressed by microglia and endothelial cells in mouse brain. A Representative immunofluorescence images of Cx3cr1 CreER-YFP -WT ( N = 4) and Cx3cr1 CreER-YFP -5xFAD ( N = 6) mouse cortex stained for microglia (GFP) and Msn. Arrow indicates microglia immunopositive for GFP and Msn. Asterisk indicates cells immunopositive for Msn only. B Representative immunofluorescence images of Cx3cr1 CreER-YFP -5xFAD ( N = 6) mouse cortex stained for amyloid-beta (Aβ), microglia (GFP), and Msn. Arrow indicates Aβ plaque as well as microglia immunopositive for GFP and Msn. C Representative immunofluorescence images of WT ( N = 3) and 5xFAD ( N = 4) mouse cortex stained for endothelial cells (CD31) and Msn. Arrow indicates endothelial cells immunopositive for CD31 and Msn. Asterisk indicates cells immunopostive for Msn only. D Representative immunofluorescence images of WT ( N = 3) and 5xFAD ( N = 4) mouse cortex stained for astrocytes (GFAP) and Msn. Arrow indicates cells immunopositive for Msn only. Asterisk indicates astroctyes immunopostive for GFAP only. Scale bar = 30µm.

    Journal: bioRxiv

    Article Title: Flow-cytometric microglial sorting coupled with quantitative proteomics identifies moesin as a highly-abundant microglial protein with relevance to Alzheimer’s disease

    doi: 10.1101/802694

    Figure Lengend Snippet: Moesin is expressed by microglia and endothelial cells in mouse brain. A Representative immunofluorescence images of Cx3cr1 CreER-YFP -WT ( N = 4) and Cx3cr1 CreER-YFP -5xFAD ( N = 6) mouse cortex stained for microglia (GFP) and Msn. Arrow indicates microglia immunopositive for GFP and Msn. Asterisk indicates cells immunopositive for Msn only. B Representative immunofluorescence images of Cx3cr1 CreER-YFP -5xFAD ( N = 6) mouse cortex stained for amyloid-beta (Aβ), microglia (GFP), and Msn. Arrow indicates Aβ plaque as well as microglia immunopositive for GFP and Msn. C Representative immunofluorescence images of WT ( N = 3) and 5xFAD ( N = 4) mouse cortex stained for endothelial cells (CD31) and Msn. Arrow indicates endothelial cells immunopositive for CD31 and Msn. Asterisk indicates cells immunopostive for Msn only. D Representative immunofluorescence images of WT ( N = 3) and 5xFAD ( N = 4) mouse cortex stained for astrocytes (GFAP) and Msn. Arrow indicates cells immunopositive for Msn only. Asterisk indicates astroctyes immunopostive for GFAP only. Scale bar = 30µm.

    Article Snippet: For immunofluorescence staining, 3-4 brain sections from each mouse were thoroughly washed to remove cryopreservative, blocked in 8% normal horse serum diluted in 1× tris buffered saline (TBS) and 0.1% Triton-X for 1 hour, and incubated with primary antibodies diluted in 1×PBS overnight (1:200 goat anti-GFP [Rockland, Cat. No. 600-101-215], 1:100 rabbit anti-Msn [Abcam, Cat. No. ab52490], 1:25 rat anti-CD31 [BD Bioscience, Cat. No. 550274].

    Techniques: Immunofluorescence, Staining

    Endothelium contributes cells to aortic calcification of Ins2 Akita/+ mice (A) Aortic expression of endothelial markers CD31 and vWF in Ins2 Akita/+ mice visualized by immunostaining. (B) Immunostaining of aortic tissues from wild type (WT) and Ins2 Akita/+ mice showed co-expression of endothelial markers CD31 (left) and vWF (right) and osteogenic markers Cbfa1 and Osterix (OSX) in the Ins2 Akita/+ mice. (C) Co-expression of CD31 and Cbfa1 in enzymatically dispersed aortic cells from WT and Ins2 Akita/+ mice , as determined by FACS. (D) Aortic expression of GFP in Tie2-Gfp tg and Ins2 Akita/+ ;Tie2-Gfp tg mice by immunostaining with anti-GFP antibodies. (E) Immunostaining of aortic tissues from Tie2-Gfp tg and Ins2 Akita/+ ;Tie2-Gfp tg mice showed co-expression of GFP with Cbfa1 and OSX in the Ins2 Akita/+ ;Tie2-Gfp tg mice. Scale bars, 100 μm. DAPI (blue) was used to visualize nuclei. Non-specific IgG control showed no staining. Vessel lumen faces upwards in the photos.

    Journal: Circulation research

    Article Title: A Role for the Endothelium in Vascular Calcification

    doi: 10.1161/CIRCRESAHA.113.301792

    Figure Lengend Snippet: Endothelium contributes cells to aortic calcification of Ins2 Akita/+ mice (A) Aortic expression of endothelial markers CD31 and vWF in Ins2 Akita/+ mice visualized by immunostaining. (B) Immunostaining of aortic tissues from wild type (WT) and Ins2 Akita/+ mice showed co-expression of endothelial markers CD31 (left) and vWF (right) and osteogenic markers Cbfa1 and Osterix (OSX) in the Ins2 Akita/+ mice. (C) Co-expression of CD31 and Cbfa1 in enzymatically dispersed aortic cells from WT and Ins2 Akita/+ mice , as determined by FACS. (D) Aortic expression of GFP in Tie2-Gfp tg and Ins2 Akita/+ ;Tie2-Gfp tg mice by immunostaining with anti-GFP antibodies. (E) Immunostaining of aortic tissues from Tie2-Gfp tg and Ins2 Akita/+ ;Tie2-Gfp tg mice showed co-expression of GFP with Cbfa1 and OSX in the Ins2 Akita/+ ;Tie2-Gfp tg mice. Scale bars, 100 μm. DAPI (blue) was used to visualize nuclei. Non-specific IgG control showed no staining. Vessel lumen faces upwards in the photos.

    Article Snippet: Furthermore, CD31 and von Willebrand factor (vWF) expression was detected deep in the calcified media , and immunostaining showed co-expression of CD31 and vWF, respectively, with osteogenic markers core binding factor alpha 1 (Cbfa1) and Osterix ( ).

    Techniques: Mouse Assay, Expressing, Immunostaining, FACS, Staining

    Depletion of MGP allows for multipotency and osteoinduction in human aortic endothelial cells (HAECs) (A) Expression of multipotent markers Sox2, Nanog and Oct3/4 in HAECs after transfection of scrambled control siRNA (SCR) or MGP siRNA only (lane 1, 2), siRNA transfection with BMP4 treatment (lane 3, 4), siRNA transfection with Noggin treatment (lane 5, 6), or siRNA transfection with BMP4 and Noggin treatment (lane 7, 8), as determined by immunoblotting. (B) Co-expression of CD31 and SSEA-3, or CD31 and SSEA-4 after transfection of scrambled control siRNA or MGP siRNA without additional treatment (left), and with BMP4 treatment (right), as determined by FACS analysis. (C) Expression of multipotent markers Sox2, Nanog and Oct3/4, osteogenic markers Cbfa1, Osterix (OSX), early SMC markers SM22α, α-SM actin (αSMA), and late SMC marker SM-myosin heavy chain (SM-MHC) (top) after transfection of scrambled control siRNA or MGP siRNA and treatment with control (C), BMP2 (B), osteogenic media (O) or both (B+O), as determined by immunoblotting. (D) Staining for alkaline phosphatase (ALP), and mineral (Alizarin Red and Von Kossa staining) in HAECS treated as described in (c). (E) Expression of multipotent markers Sox2, Nanog and Oct3/4, osteogenic markers Cbfa1, OSX, SM22α, αSMA, and SM-MHC (top) after transfection of scrambled control siRNA or MGP siRNA and treatment with control (C), BMP2 (B), high glucose medium (G) or both (B+G), as determined by immunoblotting. (F) Staining for ALP and mineral in HAECS treated as described in (E).

    Journal: Circulation research

    Article Title: A Role for the Endothelium in Vascular Calcification

    doi: 10.1161/CIRCRESAHA.113.301792

    Figure Lengend Snippet: Depletion of MGP allows for multipotency and osteoinduction in human aortic endothelial cells (HAECs) (A) Expression of multipotent markers Sox2, Nanog and Oct3/4 in HAECs after transfection of scrambled control siRNA (SCR) or MGP siRNA only (lane 1, 2), siRNA transfection with BMP4 treatment (lane 3, 4), siRNA transfection with Noggin treatment (lane 5, 6), or siRNA transfection with BMP4 and Noggin treatment (lane 7, 8), as determined by immunoblotting. (B) Co-expression of CD31 and SSEA-3, or CD31 and SSEA-4 after transfection of scrambled control siRNA or MGP siRNA without additional treatment (left), and with BMP4 treatment (right), as determined by FACS analysis. (C) Expression of multipotent markers Sox2, Nanog and Oct3/4, osteogenic markers Cbfa1, Osterix (OSX), early SMC markers SM22α, α-SM actin (αSMA), and late SMC marker SM-myosin heavy chain (SM-MHC) (top) after transfection of scrambled control siRNA or MGP siRNA and treatment with control (C), BMP2 (B), osteogenic media (O) or both (B+O), as determined by immunoblotting. (D) Staining for alkaline phosphatase (ALP), and mineral (Alizarin Red and Von Kossa staining) in HAECS treated as described in (c). (E) Expression of multipotent markers Sox2, Nanog and Oct3/4, osteogenic markers Cbfa1, OSX, SM22α, αSMA, and SM-MHC (top) after transfection of scrambled control siRNA or MGP siRNA and treatment with control (C), BMP2 (B), high glucose medium (G) or both (B+G), as determined by immunoblotting. (F) Staining for ALP and mineral in HAECS treated as described in (E).

    Article Snippet: Furthermore, CD31 and von Willebrand factor (vWF) expression was detected deep in the calcified media , and immunostaining showed co-expression of CD31 and vWF, respectively, with osteogenic markers core binding factor alpha 1 (Cbfa1) and Osterix ( ).

    Techniques: Expressing, Transfection, FACS, Marker, Staining, ALP Assay

    Multipotent marker expression in Mgp −/− and Mgp −/− ;Tie2-Gfptg endothelium (A) Aortic expression of Sox2, Nanog and Oct3/4 in Mgp +/+ and Mgp − / − mice detected by immunoblotting (left) and immunostaining (right). β-actin was used as control. (B) Aortic expression of Sox2, Nanog and Oct3/4 in Tie2-Gfp tg and Mgp − / − ;Tie2-Gfp tgmice determined by immunoblotting (left) and immunostaining (right). β-actin was used as control. (C) Co-expression of CD31 and Sox2 in enzymatically dispersed CD45-negative aortic cells from Mgp +/+ and Mgp − / − mice, as determined by FACS. Scale bars, 50 μm. Non-specific IgG control showed no staining. Vessel lumen faces upwards in the photos.

    Journal: Circulation research

    Article Title: A Role for the Endothelium in Vascular Calcification

    doi: 10.1161/CIRCRESAHA.113.301792

    Figure Lengend Snippet: Multipotent marker expression in Mgp −/− and Mgp −/− ;Tie2-Gfptg endothelium (A) Aortic expression of Sox2, Nanog and Oct3/4 in Mgp +/+ and Mgp − / − mice detected by immunoblotting (left) and immunostaining (right). β-actin was used as control. (B) Aortic expression of Sox2, Nanog and Oct3/4 in Tie2-Gfp tg and Mgp − / − ;Tie2-Gfp tgmice determined by immunoblotting (left) and immunostaining (right). β-actin was used as control. (C) Co-expression of CD31 and Sox2 in enzymatically dispersed CD45-negative aortic cells from Mgp +/+ and Mgp − / − mice, as determined by FACS. Scale bars, 50 μm. Non-specific IgG control showed no staining. Vessel lumen faces upwards in the photos.

    Article Snippet: Furthermore, CD31 and von Willebrand factor (vWF) expression was detected deep in the calcified media , and immunostaining showed co-expression of CD31 and vWF, respectively, with osteogenic markers core binding factor alpha 1 (Cbfa1) and Osterix ( ).

    Techniques: Marker, Expressing, Mouse Assay, Immunostaining, FACS, Staining

    Time course of aortic changes in Mgp −/− mouse aorta (A) Aortas were collected between postnatal day (P) 2–30 from Mgp +/+ and Mgp − / − mice as indicated, and stained with H E. (B) Time course of aortic expression in Mgp +/+ and Mgp − / − aorta (P2–30) of MGP, BMP4, EC markers VE-cadherin (VE-cad) and CD31, osteogenic markers Cbfa1 and Osterix (OSX), and multipotency markers Nanog, Sox2 and Oct3/4. The expression was compared to that on P2. Scale bars, 100 μm.

    Journal: Circulation research

    Article Title: A Role for the Endothelium in Vascular Calcification

    doi: 10.1161/CIRCRESAHA.113.301792

    Figure Lengend Snippet: Time course of aortic changes in Mgp −/− mouse aorta (A) Aortas were collected between postnatal day (P) 2–30 from Mgp +/+ and Mgp − / − mice as indicated, and stained with H E. (B) Time course of aortic expression in Mgp +/+ and Mgp − / − aorta (P2–30) of MGP, BMP4, EC markers VE-cadherin (VE-cad) and CD31, osteogenic markers Cbfa1 and Osterix (OSX), and multipotency markers Nanog, Sox2 and Oct3/4. The expression was compared to that on P2. Scale bars, 100 μm.

    Article Snippet: Furthermore, CD31 and von Willebrand factor (vWF) expression was detected deep in the calcified media , and immunostaining showed co-expression of CD31 and vWF, respectively, with osteogenic markers core binding factor alpha 1 (Cbfa1) and Osterix ( ).

    Techniques: Mouse Assay, Staining, Expressing

    Time course of aortic changes in Mgp −/− mouse aorta (A–C) Aortas were collected between postnatal day (P) 2–30 from Mgp +/+ and Mgp − / − mice as indicated. They were immunostained for (A) CD31, (B) co-stained for CD31 and Sox2, and (C) co-stained for CD31 and Osterix (OSX). DAPI (blue) was used to visualize nuclei. Scale bars, 100 μm. DAPI (blue) was used to visualize nuclei. Vessel lumen faces upwards or to the right in the photos.

    Journal: Circulation research

    Article Title: A Role for the Endothelium in Vascular Calcification

    doi: 10.1161/CIRCRESAHA.113.301792

    Figure Lengend Snippet: Time course of aortic changes in Mgp −/− mouse aorta (A–C) Aortas were collected between postnatal day (P) 2–30 from Mgp +/+ and Mgp − / − mice as indicated. They were immunostained for (A) CD31, (B) co-stained for CD31 and Sox2, and (C) co-stained for CD31 and Osterix (OSX). DAPI (blue) was used to visualize nuclei. Scale bars, 100 μm. DAPI (blue) was used to visualize nuclei. Vessel lumen faces upwards or to the right in the photos.

    Article Snippet: Furthermore, CD31 and von Willebrand factor (vWF) expression was detected deep in the calcified media , and immunostaining showed co-expression of CD31 and vWF, respectively, with osteogenic markers core binding factor alpha 1 (Cbfa1) and Osterix ( ).

    Techniques: Mouse Assay, Staining

    Endothelial origin of multipotent cells in aortas of diabetic Ins2 Akita/+ mice (A) Aortic expression of Sox2, Nanog and Oct3/4 in wild type (WT), Ins2 Akita/+ , Ins2 Akita/+ ;Tie2-Gfp tg and Ins2 Akita/+ ;Tie2-Gfp tg mice. β-actin was used as control. (B) Co-expression of CD31 with Sox2, Nanog and Oct3/4 in aortas of Ins2 Akita/+ mice detected by immunostaining. (C) Co-expression of GFP with Sox2, Nanog and Oct3/4 in aortas of Tie2-Gfp tg and Ins2 Akita/+ ;Tie2-Gfp tg mice detected by immunostaining. (D) Co-expression of CD31 and Sox2 in enzymatically dispersed CD45-negative aortic cells from WT and Ins2 Akita/+ mice , as determined by FACS. (E–G) Enhanced MGP expression limits aortic expression of Sox2, Nanog and Oct3/4 in Ins2 Akita/+ mice, as determined by (E) real-time PCR, (F) immunoblotting (β-actin was used as control), and (G) immunostaining in WT, Mgp tg/wt , Ins2 Akita/+ , and Ins2 Akita/+ ;Mgp tg/wt mice. Scale bars, 50 μm. DAPI (blue) was used to visualize nuclei. Vessel lumen faces upwards in the photos.

    Journal: Circulation research

    Article Title: A Role for the Endothelium in Vascular Calcification

    doi: 10.1161/CIRCRESAHA.113.301792

    Figure Lengend Snippet: Endothelial origin of multipotent cells in aortas of diabetic Ins2 Akita/+ mice (A) Aortic expression of Sox2, Nanog and Oct3/4 in wild type (WT), Ins2 Akita/+ , Ins2 Akita/+ ;Tie2-Gfp tg and Ins2 Akita/+ ;Tie2-Gfp tg mice. β-actin was used as control. (B) Co-expression of CD31 with Sox2, Nanog and Oct3/4 in aortas of Ins2 Akita/+ mice detected by immunostaining. (C) Co-expression of GFP with Sox2, Nanog and Oct3/4 in aortas of Tie2-Gfp tg and Ins2 Akita/+ ;Tie2-Gfp tg mice detected by immunostaining. (D) Co-expression of CD31 and Sox2 in enzymatically dispersed CD45-negative aortic cells from WT and Ins2 Akita/+ mice , as determined by FACS. (E–G) Enhanced MGP expression limits aortic expression of Sox2, Nanog and Oct3/4 in Ins2 Akita/+ mice, as determined by (E) real-time PCR, (F) immunoblotting (β-actin was used as control), and (G) immunostaining in WT, Mgp tg/wt , Ins2 Akita/+ , and Ins2 Akita/+ ;Mgp tg/wt mice. Scale bars, 50 μm. DAPI (blue) was used to visualize nuclei. Vessel lumen faces upwards in the photos.

    Article Snippet: Furthermore, CD31 and von Willebrand factor (vWF) expression was detected deep in the calcified media , and immunostaining showed co-expression of CD31 and vWF, respectively, with osteogenic markers core binding factor alpha 1 (Cbfa1) and Osterix ( ).

    Techniques: Mouse Assay, Expressing, Immunostaining, FACS, Real-time Polymerase Chain Reaction

    Endothelium contributes cells to aortic calcification of Mgp −/− mice (A) Aortic wall (confocal microscopy, top 2 panels), and aortic endothelium () from wild type ( Mgp +/+ ) and Mgp − / − mice. DIC: differential interference contrast. EM: electron microscopy. Magnification for EM, 3.7 ×10 3 . (B) Aortic expression of endothelial markers CD31, VE-cadherin (VE-cad), Flk-1 and vWF in Mgp +/+ and Mgp − / − mice determined by real-time PCR, immunoblotting and immunostaining. ***, p

    Journal: Circulation research

    Article Title: A Role for the Endothelium in Vascular Calcification

    doi: 10.1161/CIRCRESAHA.113.301792

    Figure Lengend Snippet: Endothelium contributes cells to aortic calcification of Mgp −/− mice (A) Aortic wall (confocal microscopy, top 2 panels), and aortic endothelium () from wild type ( Mgp +/+ ) and Mgp − / − mice. DIC: differential interference contrast. EM: electron microscopy. Magnification for EM, 3.7 ×10 3 . (B) Aortic expression of endothelial markers CD31, VE-cadherin (VE-cad), Flk-1 and vWF in Mgp +/+ and Mgp − / − mice determined by real-time PCR, immunoblotting and immunostaining. ***, p

    Article Snippet: Furthermore, CD31 and von Willebrand factor (vWF) expression was detected deep in the calcified media , and immunostaining showed co-expression of CD31 and vWF, respectively, with osteogenic markers core binding factor alpha 1 (Cbfa1) and Osterix ( ).

    Techniques: Mouse Assay, Confocal Microscopy, Electron Microscopy, Expressing, Real-time Polymerase Chain Reaction, Immunostaining

    Histological analysis of tissue from the pig esophagus at 2.5 months post implantation of a cellularized scaffold. ( A ) Macroscopic image of excised esophagus (proximal left, distal right). Samples of tissue were excised to include the site of surgery, monitored by endoscopy, with adjacent distal and proximal tissues for histology (box). ( B – F ) Representative images of hematoxylin and eosin ( B – E ) and Masson’s trichrome ( F – H ) stained excised esophageal tissue sections: ( C , F ) are at the center of the specimen, ( D , G ) are in the distal region at the anastomosis and ( E , H ) are from the same region of an esophagus without surgery (normal control). ( I – T ). Immunoreactivity for aSMA ( I , J ); Ki67 ( K , L ) suggests continued proliferation of mucosal and submucosal cells; presence of smooth muscle myosin heavy chain (SM-MHC) ( M , N ); CD31 ( O , P ); transgelin/SM22a ( Q , R ); and the relative absence of striated myosin heavy chain ( S , T ) in tissue at the site of surgery. Scale bars: A = 6 cm, C-H = 200 μm, I-T = 200 μm.

    Journal: Scientific Reports

    Article Title: Long-term regeneration and remodeling of the pig esophagus after circumferential resection using a retrievable synthetic scaffold carrying autologous cells

    doi: 10.1038/s41598-018-22401-x

    Figure Lengend Snippet: Histological analysis of tissue from the pig esophagus at 2.5 months post implantation of a cellularized scaffold. ( A ) Macroscopic image of excised esophagus (proximal left, distal right). Samples of tissue were excised to include the site of surgery, monitored by endoscopy, with adjacent distal and proximal tissues for histology (box). ( B – F ) Representative images of hematoxylin and eosin ( B – E ) and Masson’s trichrome ( F – H ) stained excised esophageal tissue sections: ( C , F ) are at the center of the specimen, ( D , G ) are in the distal region at the anastomosis and ( E , H ) are from the same region of an esophagus without surgery (normal control). ( I – T ). Immunoreactivity for aSMA ( I , J ); Ki67 ( K , L ) suggests continued proliferation of mucosal and submucosal cells; presence of smooth muscle myosin heavy chain (SM-MHC) ( M , N ); CD31 ( O , P ); transgelin/SM22a ( Q , R ); and the relative absence of striated myosin heavy chain ( S , T ) in tissue at the site of surgery. Scale bars: A = 6 cm, C-H = 200 μm, I-T = 200 μm.

    Article Snippet: The primary antibodies utilized were mouse anti-Ki67 (Dako, Carpinteria, CA; 1:100; antigen retrieval 30 minutes at 95 °C, Diva Decloaker, Biocare Medical), mouse anti-smooth muscle actin (SMA) (Abcam, Cambridge, MA; 1:150; antigen retrieval 16 hours at 60 °C, Diva Decloaker), and rabbit anti-CD31 (Abcam; 1:500; antigen retrieval 16 hours at 60 °C, Diva Decloaker), mouse anti-CD3 epsilon (Novus Biologics, 1 microgram/ml, antigen retrieval 20 min at 95 °C), rabbit anti-SM22a (abcam, 1:100), and mouse anti-SM-MHC/MYH11 (smooth muscle myosin heavy chain; MyBioSource, neat, antigen retrieval 20 minutes at 95 °C).

    Techniques: Staining

    Histological characterization of the esophageal submucosa and regenerated mucosa. Representative immunohistochemical analysis demonstrates immunoreactivity for Ki67 ( A ) suggesting continued proliferation of mucosal and submucosal cells, CD31 ( B ), CD3e ( C ), aSMA ( D ), transgelin/SM22a ( E ) and a relative absence of striated myosin heavy chain ( F ) in tissue at the site of surgery. Scale bars = 50 μm.

    Journal: Scientific Reports

    Article Title: Long-term regeneration and remodeling of the pig esophagus after circumferential resection using a retrievable synthetic scaffold carrying autologous cells

    doi: 10.1038/s41598-018-22401-x

    Figure Lengend Snippet: Histological characterization of the esophageal submucosa and regenerated mucosa. Representative immunohistochemical analysis demonstrates immunoreactivity for Ki67 ( A ) suggesting continued proliferation of mucosal and submucosal cells, CD31 ( B ), CD3e ( C ), aSMA ( D ), transgelin/SM22a ( E ) and a relative absence of striated myosin heavy chain ( F ) in tissue at the site of surgery. Scale bars = 50 μm.

    Article Snippet: The primary antibodies utilized were mouse anti-Ki67 (Dako, Carpinteria, CA; 1:100; antigen retrieval 30 minutes at 95 °C, Diva Decloaker, Biocare Medical), mouse anti-smooth muscle actin (SMA) (Abcam, Cambridge, MA; 1:150; antigen retrieval 16 hours at 60 °C, Diva Decloaker), and rabbit anti-CD31 (Abcam; 1:500; antigen retrieval 16 hours at 60 °C, Diva Decloaker), mouse anti-CD3 epsilon (Novus Biologics, 1 microgram/ml, antigen retrieval 20 min at 95 °C), rabbit anti-SM22a (abcam, 1:100), and mouse anti-SM-MHC/MYH11 (smooth muscle myosin heavy chain; MyBioSource, neat, antigen retrieval 20 minutes at 95 °C).

    Techniques: Immunohistochemistry

    Vascularized TOC devices permit robust growth of colorectal and pancreatic cancer cells. Three colorectal cancer (CRC268, CRC663, and CRC1180) and two pancreatic ductal adenocarcinoma cell lines (PDAC162 and PDAC175) were introduced into the side chambers with pre-formed vasculature in the central chamber. A: Representative immune fluorescence images taken at Days 2 and 9 after tumor cells were introduced. Green: GFP labeled endothelial cells formed vasculature. Red: RFP labelled tumor cells. The white dotted lines indicate the top and bottom boundaries of the central chamber. B: Quantification of angiogenesis, tumor cell proliferation, and migration/invasion on Day 9. Eight devices per tumor cell line were used for quantification. C: IHC images on parental CRC and PDAC tumor sections stained with anti-CD31. Representative images are shown at 100x and 200x magnification. D: Quantification of CD31-positive areas in parental tumors using IHC images. * p

    Journal: bioRxiv

    Article Title: Tumor-on-a-chip platform to interrogate the role of macrophages in tumor progression

    doi: 10.1101/2020.05.27.119636

    Figure Lengend Snippet: Vascularized TOC devices permit robust growth of colorectal and pancreatic cancer cells. Three colorectal cancer (CRC268, CRC663, and CRC1180) and two pancreatic ductal adenocarcinoma cell lines (PDAC162 and PDAC175) were introduced into the side chambers with pre-formed vasculature in the central chamber. A: Representative immune fluorescence images taken at Days 2 and 9 after tumor cells were introduced. Green: GFP labeled endothelial cells formed vasculature. Red: RFP labelled tumor cells. The white dotted lines indicate the top and bottom boundaries of the central chamber. B: Quantification of angiogenesis, tumor cell proliferation, and migration/invasion on Day 9. Eight devices per tumor cell line were used for quantification. C: IHC images on parental CRC and PDAC tumor sections stained with anti-CD31. Representative images are shown at 100x and 200x magnification. D: Quantification of CD31-positive areas in parental tumors using IHC images. * p

    Article Snippet: Anti-CD31 (#JC70A, Agilent, Santa Clara, CA) was used to stain tissue sections.

    Techniques: Fluorescence, Labeling, Migration, Immunohistochemistry, Staining

    Surface markers of synovial MSCs before and 48 h after preservation. a Representative histogram of synovial MSCs by CD44-PE-Cy7, CD73-V450, CD90-PE, CD105-APC, and CD34-PE-Cy5. b Representative dot plot of synovial MSCs by CD31-FITC and CD45-APC-H7 as negative markers. c Calculated live cell number per tube (2 million synovial MSCs). Median values and interquartile ranges are shown ( n = 4). ND not detected

    Journal: Stem Cell Research & Therapy

    Article Title: Complete human serum maintains viability and chondrogenic potential of human synovial stem cells: suitable conditions for transplantation

    doi: 10.1186/s13287-017-0596-0

    Figure Lengend Snippet: Surface markers of synovial MSCs before and 48 h after preservation. a Representative histogram of synovial MSCs by CD44-PE-Cy7, CD73-V450, CD90-PE, CD105-APC, and CD34-PE-Cy5. b Representative dot plot of synovial MSCs by CD31-FITC and CD45-APC-H7 as negative markers. c Calculated live cell number per tube (2 million synovial MSCs). Median values and interquartile ranges are shown ( n = 4). ND not detected

    Article Snippet: The cells before and 48 h after preservation were suspended in Hank’s balanced salt solution (HBSS) at a density of 5 × 105 cells/mL and stained for 30 min on ice with the antibodies CD44-PE-Cy7, CD73-V450, CD90-PE, CD105-APC, CD34-PE-Cy5, CD45-APC-H7, and CD31-FITC (all from BD), and Ghost Dye Violet 510 for dead cells (Tonbo Biosciences, CA, USA).

    Techniques: Preserving

    Characterization of the P6C cells. (A) Holoclone formation of the P6C cells. Cells from primary colorectal cancer tissue and paired normal colon tissue were trypsinized and seeded into a 6-well plate. A primary clone from a single cancer cell and a holoclone of the P6C cells (passage 20) are shown in the top column, respectively. A representative primary sphere from the colon cancer tissue and dispersed normal colon cells are shown in the bottom column. Scale bars, 200 μm. (B) Expression of distinct markers in the P6C cells. Suspended P6C cells were incubated with anti-CD45-FITC, CD31-FITC, and CD24-FITC, respectively, and were analysed by flow cytometry. Donkey anti-mouse-FITC was used as a control. M1, negative; M2, positive. (C) Surface expression of CD44 in P6C cells as detected by immunofluorescence. P6C cells were grown attached to plates for 5 d, fixed with paraformaldehyde and incubated with anti-CD44 antibody. DAPI was used to stain the nucleus. Scale bar, 100 μm. (D) Relative expression levels of distinct markers under different culture conditions, including a P6C sphere, a P6C clone and SW480 cells. Suspended cells were collected and incubated with FITC conjugated antibodies and were analysed by flow cytometry. Each sample was analysed in triplicate, and the experiment was repeated 3 times. b P

    Journal: Acta Pharmacologica Sinica

    Article Title: Establishment of a human colorectal cancer cell line P6C with stem cell properties and resistance to chemotherapeutic drugs

    doi: 10.1038/aps.2013.56

    Figure Lengend Snippet: Characterization of the P6C cells. (A) Holoclone formation of the P6C cells. Cells from primary colorectal cancer tissue and paired normal colon tissue were trypsinized and seeded into a 6-well plate. A primary clone from a single cancer cell and a holoclone of the P6C cells (passage 20) are shown in the top column, respectively. A representative primary sphere from the colon cancer tissue and dispersed normal colon cells are shown in the bottom column. Scale bars, 200 μm. (B) Expression of distinct markers in the P6C cells. Suspended P6C cells were incubated with anti-CD45-FITC, CD31-FITC, and CD24-FITC, respectively, and were analysed by flow cytometry. Donkey anti-mouse-FITC was used as a control. M1, negative; M2, positive. (C) Surface expression of CD44 in P6C cells as detected by immunofluorescence. P6C cells were grown attached to plates for 5 d, fixed with paraformaldehyde and incubated with anti-CD44 antibody. DAPI was used to stain the nucleus. Scale bar, 100 μm. (D) Relative expression levels of distinct markers under different culture conditions, including a P6C sphere, a P6C clone and SW480 cells. Suspended cells were collected and incubated with FITC conjugated antibodies and were analysed by flow cytometry. Each sample was analysed in triplicate, and the experiment was repeated 3 times. b P

    Article Snippet: Supplementary Information Controls of anti-CD45, anti-CD31 and anti-CD24 antibodies.

    Techniques: Expressing, Incubation, Flow Cytometry, Cytometry, Immunofluorescence, Staining

    Histopathology analysis of tumors reveals antibody-labeled nanoparticles localize to regions rich with immune cells. ( A ) Analysis of Prussian blue–positive (nanoparticle-rich) areas of tumors from nude mice injected with BH nanoparticles reveals only weak correlation with HER2 expression. ( B ) Conversely, this correlation is stronger in tumors from NSG mice. ( C and D ) Weak or no correlation was observed between BH nanoparticle presence and CD31 + (vascular endothelium) regions. ( E ) Representative histology images of sequential sections showing IBA-1 + cells associated with Prussian blue–positive areas in HCC1954 (HER2 + ) tumors grown in NSG mice and treated with BH (a) hematoxylin and eosin (H E), (b) Prussian blue, (c) HER2 IHC, (d) IBA-1 IHC, (e) CD-31 IHC, (f) H E of another area from same tumor, (g) sequential section stained for Prussian blue shows positive staining for iron nanoparticles, and (h) immunofluorescence (IF) staining for IBA-1 shows positivity in the nanoparticle accumulated region. ( F and G ) Iron recovery from HER2 + (HCC1954) or HER2 − (MDA-MB-231) tumors is similar whether BNF nanoparticles have trastuzumab (anti-HER2) or human IgG (polyclonal), suggesting that antibody-antigen binding does not drive intratumor nanoparticle accumulation. ns, not statistically significant.

    Journal: Science Advances

    Article Title: Nanoparticle interactions with immune cells dominate tumor retention and induce T cell–mediated tumor suppression in models of breast cancer

    doi: 10.1126/sciadv.aay1601

    Figure Lengend Snippet: Histopathology analysis of tumors reveals antibody-labeled nanoparticles localize to regions rich with immune cells. ( A ) Analysis of Prussian blue–positive (nanoparticle-rich) areas of tumors from nude mice injected with BH nanoparticles reveals only weak correlation with HER2 expression. ( B ) Conversely, this correlation is stronger in tumors from NSG mice. ( C and D ) Weak or no correlation was observed between BH nanoparticle presence and CD31 + (vascular endothelium) regions. ( E ) Representative histology images of sequential sections showing IBA-1 + cells associated with Prussian blue–positive areas in HCC1954 (HER2 + ) tumors grown in NSG mice and treated with BH (a) hematoxylin and eosin (H E), (b) Prussian blue, (c) HER2 IHC, (d) IBA-1 IHC, (e) CD-31 IHC, (f) H E of another area from same tumor, (g) sequential section stained for Prussian blue shows positive staining for iron nanoparticles, and (h) immunofluorescence (IF) staining for IBA-1 shows positivity in the nanoparticle accumulated region. ( F and G ) Iron recovery from HER2 + (HCC1954) or HER2 − (MDA-MB-231) tumors is similar whether BNF nanoparticles have trastuzumab (anti-HER2) or human IgG (polyclonal), suggesting that antibody-antigen binding does not drive intratumor nanoparticle accumulation. ns, not statistically significant.

    Article Snippet: For CD31 (Dianova, DIA 310), and IBA-1 (Wako, 019-19741), after deparaffinization and hydration, the slides were steamed in HTTR or EDTA buffer for 45 min in a steamer followed by washing in PBS containing Tween.

    Techniques: Histopathology, Labeling, Mouse Assay, Injection, Expressing, Immunohistochemistry, Staining, Immunofluorescence, Multiple Displacement Amplification, Binding Assay

    Myocardial histology. ( A ) Representative image of histology sampling from infarcted areas ( B , C , E ); peri-infarcted areas ( D , F , G ); ( B ) Masson’s Trichrome ×1.25 ( C ) Hematoxylin-eosin ×5 ( D ) Integrin α v β 3 ×20 ( E ) α-smooth muscle actin ×20 ( F ) CD68 ×20 ( G ) CD31 ×20 ( H ) Percentage fibrosis in all sample areas (I) α-smooth muscle actin percentage in scar ( J ) CD31 vessel density per high power field. pASC: porcine adipose-derived stromal cells in alginate hydrogel, Arrows show co-localization of integrin α v β 3 and CD31.

    Journal: Diagnostics

    Article Title: Angiogenesis PET Tracer Uptake (68Ga-NODAGA-E[(cRGDyK)]2) in Induced Myocardial Infarction and Stromal Cell Treatment in Minipigs

    doi: 10.3390/diagnostics8020033

    Figure Lengend Snippet: Myocardial histology. ( A ) Representative image of histology sampling from infarcted areas ( B , C , E ); peri-infarcted areas ( D , F , G ); ( B ) Masson’s Trichrome ×1.25 ( C ) Hematoxylin-eosin ×5 ( D ) Integrin α v β 3 ×20 ( E ) α-smooth muscle actin ×20 ( F ) CD68 ×20 ( G ) CD31 ×20 ( H ) Percentage fibrosis in all sample areas (I) α-smooth muscle actin percentage in scar ( J ) CD31 vessel density per high power field. pASC: porcine adipose-derived stromal cells in alginate hydrogel, Arrows show co-localization of integrin α v β 3 and CD31.

    Article Snippet: Furthermore, we found no difference in SUVindex between pASC-treated vs. non-pASC-treated minipigs, which also was reflected in the histological findings of similar CD31 vessel density in the groups, with co-localization of integrin αv β3 and CD31.

    Techniques: Sampling, Derivative Assay

    Keratin, Ezh2 and CD31 expression in tumors treated with SKI-606 for 14 days. Luminal epithelial lineages (K8, red) and basal-like epithelia (K14, green) were visualized in sections of tumors of mice treated with vehicle (A-a) or SKI606. (A-b). B, localization of Ezh2 and E-cadherin in tumors. Lower panels show decreased Ezh2 expression of SKI-606-treated tumors. C, CD31 (red) staining of tumor sections. SKI-606-treated tumor (right) has more organized and regular spacing. D, image analysis reveals length of vessel perimeter (left), endothelium area of vessel (middle), by t-test and vessel density (right). *P

    Journal: Oncogene

    Article Title: Control of mammary tumor differentiation by SKI-606 (bosutinib)

    doi: 10.1038/onc.2010.412

    Figure Lengend Snippet: Keratin, Ezh2 and CD31 expression in tumors treated with SKI-606 for 14 days. Luminal epithelial lineages (K8, red) and basal-like epithelia (K14, green) were visualized in sections of tumors of mice treated with vehicle (A-a) or SKI606. (A-b). B, localization of Ezh2 and E-cadherin in tumors. Lower panels show decreased Ezh2 expression of SKI-606-treated tumors. C, CD31 (red) staining of tumor sections. SKI-606-treated tumor (right) has more organized and regular spacing. D, image analysis reveals length of vessel perimeter (left), endothelium area of vessel (middle), by t-test and vessel density (right). *P

    Article Snippet: K8, K14, smooth muscle actin and CD31 were detected with acetone-fixed, sucrose infused, OCT embedded, frozen sections.

    Techniques: Expressing, Mouse Assay, Staining