rat anti-cd31 Search Results


  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 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 2640 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rat anti mouse cd31/product/Becton Dickinson
    Average 93 stars, based on 2640 article reviews
    Price from $9.99 to $1999.99
    rat anti mouse cd31 - by Bioz Stars, 2020-09
    93/100 stars
      Buy from Supplier

    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 1729 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rat anti cd31/product/Becton Dickinson
    Average 94 stars, based on 1729 article reviews
    Price from $9.99 to $1999.99
    rat anti cd31 - by Bioz Stars, 2020-09
    94/100 stars
      Buy from Supplier

    99
    R&D Systems mouse rat cd31 pecam 1 antibody
    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.
    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
    https://www.bioz.com/result/mouse rat cd31 pecam 1 antibody/product/R&D Systems
    Average 99 stars, based on 554 article reviews
    Price from $9.99 to $1999.99
    mouse rat cd31 pecam 1 antibody - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    92
    Pharmingen rat anti mouse cd31
    Assessment of tumor malignancy based on <t>CD31,</t> MMP13 and TGF-β1 expression. (A) H E staining of sliced tissue samples from Non-Rec and Rec tumors. Areas containing angiogenic features are demarcated by the red lines and indicated with yellow arrows. The yellow arrow-heads indicated the elongated fibroblast. An enlarged view of the angiogenic area is shown to the right of each sample. (B) Immunofluorescent staining of Non-Rec and Rec tumor tissue samples for CD31 and MMP13. Colocalized staining of CD31 and MMP13 is indicated by the white arrows. (C) The bar graph demonstrates the difference in microvascular density (MVD) between the groups. (D) The ELISA assay showed that TGF-β1 was expressed in a recurrence-status-dependent manner, because the level of released TGF-β1 was lower in the culture medium from the Non-Rec group than from the Rec group. Scale bar = 100 μm in the H E images; Scale bar = 200 μm in the Immunofluorescent images. *, p
    Rat Anti Mouse Cd31, supplied by Pharmingen, used in various techniques. Bioz Stars score: 92/100, based on 275 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rat anti mouse cd31/product/Pharmingen
    Average 92 stars, based on 275 article reviews
    Price from $9.99 to $1999.99
    rat anti mouse cd31 - by Bioz Stars, 2020-09
    92/100 stars
      Buy from Supplier

    91
    Becton Dickinson rat monoclonal anti cd31
    Tumor vessel perfusion. The vascular function is increased in tumors of RT pos SG ko versus RT pos SG wt mice. 15 week RT pos SG wt (a) and RT pos SG ko (b) mice were perfused with FITC-lectin, which was introduced to the circulation prior to sacrifice of the animals and excision of the pancreas. Tumor sections were stained for the endothelial marker <t>CD31</t> (red), the localization of which was compared to lectin (green) that had immobilized on the luminal side of endothelial cells in perfused vessels. The ratio between immobilized lectin and CD31 staining was calculated to determine the fraction of perfused, and thus functional vessels (c). Each data point in represents an individual animal. Statistical analysis was performed using a two-tailed Mann-Whitney test. Error bars represent mean ± SEM.
    Rat Monoclonal Anti Cd31, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 91/100, based on 262 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rat monoclonal anti cd31/product/Becton Dickinson
    Average 91 stars, based on 262 article reviews
    Price from $9.99 to $1999.99
    rat monoclonal anti cd31 - by Bioz Stars, 2020-09
    91/100 stars
      Buy from Supplier

    92
    Dianova rat anti mouse cd31
    Loss of Vav3 results in microvascular leakage in vivo. (A) Enhanced extravasation of red fluorescent microspheres in the trachea of Vav3 -deficient mice. Control and Vav3 −/− mice were injected (i.v.) with 23-nm red fluorescent microspheres (red) in combination with VEGF (3 μg per animal). Whole-mount preparations of trachea were immunostained with <t>anti-CD31</t> (green). Bottom panels show magnification of boxed area in top panels (bars, 50 µm). (B) Quantification of microvascular leakage by determining area of microsphere extravasation versus total vessel area. Percentage of microsphere extravasation is set to 100% for trachea WT controls compared with Vav3 −/− (mean ± SEM; *, P
    Rat Anti Mouse Cd31, supplied by Dianova, used in various techniques. Bioz Stars score: 92/100, based on 140 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rat anti mouse cd31/product/Dianova
    Average 92 stars, based on 140 article reviews
    Price from $9.99 to $1999.99
    rat anti mouse cd31 - by Bioz Stars, 2020-09
    92/100 stars
      Buy from Supplier

    92
    Thermo Fisher rat anti cd31
    Whole mount imaging of meningeal lymphatic vessels in mice. (a) Meningeal LVs were characterized by whole-mount immunofluorescence staining for <t>CD31</t> (green) and Lyve-1 (red). Low magnification view of meningeal LVs. LVs run alongside the sagittal sinus (ss) and transverse sinus (ts). The boxed regions in panel A outline the areas that we analyzed in our stroke experiments to acquire lymphatic vessel index (LVI) at the ss. (b) Representative images showing Prox1 (red) and podoplanin (green) expression by meningeal LVs around the transverse sinus (TS). (c) A 3D rendering of meningeal LVs (red) in a Prox1-tdTomato transgenic mouse was generated using serial two-photon tomography. The signal from the hippocampus was digitally removed. (a), scale bar is 1000 µm; (b), scale bar is 20 µm.
    Rat Anti Cd31, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 92/100, based on 164 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rat anti cd31/product/Thermo Fisher
    Average 92 stars, based on 164 article reviews
    Price from $9.99 to $1999.99
    rat anti cd31 - by Bioz Stars, 2020-09
    92/100 stars
      Buy from Supplier

    92
    Pharmingen rat anti cd31
    PLX4720 treatment stabilizes blood vessels in COLO205 and SK-MEL-28 xenograft models. ( A ) Representative images of immunofluorescence analysis with the <t>CD31</t> endothelial marker (green) in COLO205 ( a – c ) and SK-MEL-28 ( d – f ) tumors. (Scale
    Rat Anti Cd31, supplied by Pharmingen, used in various techniques. Bioz Stars score: 92/100, based on 112 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rat anti cd31/product/Pharmingen
    Average 92 stars, based on 112 article reviews
    Price from $9.99 to $1999.99
    rat anti cd31 - by Bioz Stars, 2020-09
    92/100 stars
      Buy from Supplier

    88
    Becton Dickinson rat anti mouse cd31 pecam 1 monoclonal antibody
    PLX4720 treatment stabilizes blood vessels in COLO205 and SK-MEL-28 xenograft models. ( A ) Representative images of immunofluorescence analysis with the <t>CD31</t> endothelial marker (green) in COLO205 ( a – c ) and SK-MEL-28 ( d – f ) tumors. (Scale
    Rat Anti Mouse Cd31 Pecam 1 Monoclonal Antibody, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 88/100, based on 89 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rat anti mouse cd31 pecam 1 monoclonal antibody/product/Becton Dickinson
    Average 88 stars, based on 89 article reviews
    Price from $9.99 to $1999.99
    rat anti mouse cd31 pecam 1 monoclonal antibody - by Bioz Stars, 2020-09
    88/100 stars
      Buy from Supplier

    92
    Dianova rat anti cd31
    F8-VEGFC induces expansion of the lymphatic vasculature in chronic colitis. (A) Representative pictures of immunofluorescence labeling of the inflamed colon after F8-SIP (upper panels) or F8-VEGFC (lower panels) treatment for LYVE-1 (green), <t>CD31</t> (red), and Hoechst (blue). Dotted white lines mark the separation between the mucosa (M) and the submucosa (SM). Scale bars: 100 μm. LYVE-1 + LVs were quantified in terms of number (B) and percentage of stained area (C) in 5–8 fields per colon per mouse ( n = 5 per group). CD31+LYVE-1- blood vessels were quantified in terms of number (D) and percentage of stained area (E) in 5–8 fields per colon per mouse ( n = 5 per group). All data are presented as mean ± SD. Statistical significance was determined by two-tailed Student’s t test. Asterisks indicate statistical significance with p
    Rat Anti Cd31, supplied by Dianova, used in various techniques. Bioz Stars score: 92/100, based on 104 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rat anti cd31/product/Dianova
    Average 92 stars, based on 104 article reviews
    Price from $9.99 to $1999.99
    rat anti cd31 - by Bioz Stars, 2020-09
    92/100 stars
      Buy from Supplier

    92
    Thermo Fisher rat anti mouse cd31
    A model for the <t>CD31-induced</t> barrier response. MHC triggering induces RhoA and Erk activation and EC contraction (1). Erk phosphorylation is modulated by CD31 signals, possibly via SHP-2 (2). MHC signals induce CD31 ITIM phosphorylation and SHP-2 recruitment. SHP-2 prevents the phosphorylation of b-catenin (5) and VE-cadherin (6), thus stabilizing the junctional complex. In addition, dephosphorylated b-catenin can transfer to the nucleus where it induces cMyc transcription. In parallel, SHP-2 induces AKT activation which in turn inhibits FoxO1 nuclear translocation, thus preventing inhibition of cMyc transcription. This leads to enhanced transcription of glycolysis enzymes and enhanced glycolysis required for actin remodeling and maintenance of junctional anchorage.
    Rat Anti Mouse Cd31, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 92/100, based on 149 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rat anti mouse cd31/product/Thermo Fisher
    Average 92 stars, based on 149 article reviews
    Price from $9.99 to $1999.99
    rat anti mouse cd31 - by Bioz Stars, 2020-09
    92/100 stars
      Buy from Supplier

    96
    Bio-Rad rat anti mouse cd31
    Skin phenotype of Aspn -/- mice. (A) Asporin immunoblotting of dorsal skin extracts from three wild-type and three Aspn -/- mice. (B) 2 month-old female dorsal skins were analyzed histologically by staining with Massson Trichrome, or (C) by immunohistochemistry by staining for the presence of <t>CD31-positive</t> blood vessels and F4/80-positive macrophages; bars in A and B are 100 μm. Similar results were obtained from analyses of six mice from each genotype. (D) Transmission electron microscopy on cross-sectioned collagen fibrils in reticular dermis. (E) Transmission electron microscopy on longitudinally oriented collagen fibrils in reticular dermis. Bars in C and D are 200 nm. (F) Quantification of collagen fibril diameter in reticular dermis. 1,000 fibrils were measured from electron microscopy images collected from six wild-type and six Aspn -/- mice.
    Rat Anti Mouse Cd31, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 96/100, based on 96 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rat anti mouse cd31/product/Bio-Rad
    Average 96 stars, based on 96 article reviews
    Price from $9.99 to $1999.99
    rat anti mouse cd31 - by Bioz Stars, 2020-09
    96/100 stars
      Buy from Supplier

    99
    Abcam rat anti cd31
    Overexpression of miR‐21 abrogates dihydromyricetin (DMY)‐increased DDAH1 expression and endothelial nitric oxide (NO) synthase phosphorylation and reduced ADMA levels. HUVECs were transfected with 100 n m agomir NC or miR‐21 agomir for 24 h and then treated with ox‐LDL (120 μg/mL) or ox‐LDL plus DMY (25 μmol/L) for 16 h and harvested for indicated experiments. A Western blot analysis of DDAH1 and p‐endothelial NO synthase (eNOS; ser1177) expression. n = 3 independent experiments. B, ELISA analysis of ADMA levels in cell lysates. n = 3 independent experiment. C, D, Representative images and quantification show DDAH1 (C) and phosphor‐eNOS (ser1177) (D) expression in endothelial cells in the aortic sinus lesions from DMY combined with NS‐m injection or DMY combined with 21‐m injection‐treated Apoe − / − mice fed with HFD for 12 wk. Frozen sections of aortic sinus were stained for anti‐DDAH1 or p‐eNOS (green), <t>anti‐CD31</t> (red) and DAPI (blue). The dashed line area indicates differential DDAH1 or phosphor‐eNOS expression in endothelial cells. Scale: 50 μm. n = 7 mice per group. E, Western blot analysis of DDAH1 and p‐eNOS (ser1177) expression in liver. n = 7 mice per group. F and G, ELISA analysis of ADMA levels in plasma (F) and livers (G) from DMY combined with NS‐m injection or DMY combined with 21‐m injection‐treated Apoe − / − mice fed with HFD for 12 wk. n = 7 mice per group. Data shown are mean ± SEM. * P
    Rat Anti Cd31, supplied by Abcam, used in various techniques. Bioz Stars score: 99/100, based on 65 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rat anti cd31/product/Abcam
    Average 99 stars, based on 65 article reviews
    Price from $9.99 to $1999.99
    rat anti cd31 - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    94
    Bio-Rad cd31
    Effect of AMI on rat SVF cells. a Cell size (μm) of the SVF cells, demonstrating no difference between the three groups. b Percentage of colony-forming cells, showing significantly fewer colonies in the 1D group (6.1 ± 1.6 %) compared to the Control (11.4 ± 1.8 %) and 7D groups (11.0 ± 0.9 %). c Number of cells, obtained 4 days after seeding of the SVF fraction, demonstrating significantly fewer cells in the 1D group (1.3 ± 0.1 million cells) compared to the Control (2.0 ± 0.1 million cells) and 7D groups (1.8 ± 0.1 million cells). d Percentage of positive cells for the markers <t>CD31,</t> CD34, CD45, CD73, CD90, CD105 and CD271 showing significantly fewer CD90- and CD105-positive cells in the 1D group. Data shown as mean ± SD (Control and 7D group n = 6, 1D group n = 5, * p
    Cd31, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 94/100, based on 709 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/cd31/product/Bio-Rad
    Average 94 stars, based on 709 article reviews
    Price from $9.99 to $1999.99
    cd31 - by Bioz Stars, 2020-09
    94/100 stars
      Buy from Supplier

    Image Search Results


    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

    Effect of alcohol exposure on VEGF expression and angiogenesis in mice. (A) E0771 mouse breast cancer cells were implanted in mammary fat pads of C57BL6 mice. The mice were exposed to alcohol in drinking water for two weeks. Mice were sacrificed and mammary tumors were dissected and sectioned for VEGF IHC as described in Materials and methods. (B) Tumor microvessels were identified by CD31 IHC and the AMVD was quantified and expressed as the number of microvessels/mm 2 area. The results are represented as the mean ± SEM of 20–22 animals. * Denotes a statistically significant difference (P

    Journal: Oncology Letters

    Article Title: Alcohol promotes mammary tumor growth through activation of VEGF-dependent tumor angiogenesis

    doi: 10.3892/ol.2014.2146

    Figure Lengend Snippet: Effect of alcohol exposure on VEGF expression and angiogenesis in mice. (A) E0771 mouse breast cancer cells were implanted in mammary fat pads of C57BL6 mice. The mice were exposed to alcohol in drinking water for two weeks. Mice were sacrificed and mammary tumors were dissected and sectioned for VEGF IHC as described in Materials and methods. (B) Tumor microvessels were identified by CD31 IHC and the AMVD was quantified and expressed as the number of microvessels/mm 2 area. The results are represented as the mean ± SEM of 20–22 animals. * Denotes a statistically significant difference (P

    Article Snippet: Rat anti-mouse CD31 monoclonal antibody was obtained from BD Biosciences (San Diego, CA, US), while anti-VEGF antibody was obtained from Santa Cruz Biotechnology, Inc. (Dallas, TX, USA).

    Techniques: Expressing, Mouse Assay, Immunohistochemistry

    Immunohistochemical analyses of OSC-19 xenograft tumors in nude mice. A, Tumors were harvested after 7 d of treatment, and representative sections obtained from OSC-19 tumors were immunostained for expression of CD31 (endothelial cell marker) and TUNEL

    Journal: Clinical cancer research : an official journal of the American Association for Cancer Research

    Article Title: Vandetanib Restores Head and Neck Squamous Cell Carcinoma Cells' Sensitivity to Cisplatin and Radiation In Vivo and In Vitro

    doi: 10.1158/1078-0432.CCR-10-2120

    Figure Lengend Snippet: Immunohistochemical analyses of OSC-19 xenograft tumors in nude mice. A, Tumors were harvested after 7 d of treatment, and representative sections obtained from OSC-19 tumors were immunostained for expression of CD31 (endothelial cell marker) and TUNEL

    Article Snippet: Rat monoclonal anti-mouse CD31 (platelet-endothelial cell adhesion molecule 1; PECAM), the primary antibody for immunohistochemical analysis, was purchased from BD Pharmingen (San Diego, CA).

    Techniques: Immunohistochemistry, Mouse Assay, Expressing, Marker, TUNEL Assay

    Immunofluorescence TF/CD31 double-staining of MDA-MB-231 tumor and MDA-MB-435 tumor. Scale bar: 100 μm.

    Journal: European journal of nuclear medicine and molecular imaging

    Article Title: ImmunoPET of Tissue Factor Expression in Triple-Negative Breast Cancer with a Radiolabeled Antibody Fab Fragment

    doi: 10.1007/s00259-015-3038-1

    Figure Lengend Snippet: Immunofluorescence TF/CD31 double-staining of MDA-MB-231 tumor and MDA-MB-435 tumor. Scale bar: 100 μm.

    Article Snippet: Rat anti-mouse CD31 primary antibody was purchased from BD Biosciences (San Diego, CA).

    Techniques: Immunofluorescence, Double Staining, Multiple Displacement Amplification

    Analysis of GLV-1h68 virus-induced changes in HuH7- or PLC-tumor vascularization by confocal laser microscopy. Determination of vascular density using CD31 immunohistochemistry in virus- treated and non-treated HuH7 or PLC, tumors (A). The vascular density was measured in CD31-labeled tumor cross-sections (n = 6 per group) and presented as mean values +/− standard deviations. The asterisks (**) indicate a significant difference between experimental groups (** P

    Journal: PLoS ONE

    Article Title: Efficient Colonization and Therapy of Human Hepatocellular Carcinoma (HCC) Using the Oncolytic Vaccinia Virus Strain GLV-1h68

    doi: 10.1371/journal.pone.0022069

    Figure Lengend Snippet: Analysis of GLV-1h68 virus-induced changes in HuH7- or PLC-tumor vascularization by confocal laser microscopy. Determination of vascular density using CD31 immunohistochemistry in virus- treated and non-treated HuH7 or PLC, tumors (A). The vascular density was measured in CD31-labeled tumor cross-sections (n = 6 per group) and presented as mean values +/− standard deviations. The asterisks (**) indicate a significant difference between experimental groups (** P

    Article Snippet: Endothelial cells were labeled with monoclonal rat anti-mouse CD31 antibody (BD Pharmingen, San Diego, CA) or hamster anti-mouse CD31 antibody (Chemicon, International, Temecula, CA).

    Techniques: Planar Chromatography, Microscopy, Immunohistochemistry, Labeling

    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

    Cardiovascular abnormalities in homozygous Rasa1 R780Q mice. A and B: Whole mount ( A ) and sections ( B ) of E9.5 Rasa1 R780Q/R780Q and Rasa1 fl/fl embryos were stained with an anti-CD31 antibody to reveal blood vasculature. A: Note the organized BV network in the Rasa1 fl/fl yolk sac (YS) and honeycombed appearance of blood vasculature in the Rasa1 R780Q/R780Q yolk sac ( asterisk ). Note also irregular blood vasculature in the Rasa1 R780Q/R780Q embryo proper (E). B: Note the disorganized dorsal aorta in the Rasa1 R780Q/R780Q embryo compared with the Rasa1 fl/fl embryo ( arrows ). DA, dorsal aorta; IS, intersegmental artery.

    Journal: The American Journal of Pathology

    Article Title: Blood Vascular Abnormalities in Rasa1R780Q Knockin Mice

    doi: 10.1016/j.ajpath.2014.08.018

    Figure Lengend Snippet: Cardiovascular abnormalities in homozygous Rasa1 R780Q mice. A and B: Whole mount ( A ) and sections ( B ) of E9.5 Rasa1 R780Q/R780Q and Rasa1 fl/fl embryos were stained with an anti-CD31 antibody to reveal blood vasculature. A: Note the organized BV network in the Rasa1 fl/fl yolk sac (YS) and honeycombed appearance of blood vasculature in the Rasa1 R780Q/R780Q yolk sac ( asterisk ). Note also irregular blood vasculature in the Rasa1 R780Q/R780Q embryo proper (E). B: Note the disorganized dorsal aorta in the Rasa1 R780Q/R780Q embryo compared with the Rasa1 fl/fl embryo ( arrows ). DA, dorsal aorta; IS, intersegmental artery.

    Article Snippet: Fixed embryos were stained with a rat anti-CD31 antibody (BD Biosciences, San Jose, CA), followed by a secondary goat anti-rat Ig Alexa Fluor 488 antibody (Invitrogen, Carlsbad, CA).

    Techniques: Mouse Assay, Staining

    MAPK activation in homozygous Rasa1 R780Q embryos. Tissue sections of E9.5 Rasa1 R780Q/R780Q and Rasa1 fl/fl embryos were stained with anti-CD31 (red) and anti–phospho-ERK (green) antibodies. Shown are select regions of embryos that include part of the dorsal aorta ( arrows ). Note the absence of pERK staining in most ECs in Rasa1 fl/fl embryos and the presence of pERK staining in most ECs in Rasa1 R780Q/R780Q embryos.

    Journal: The American Journal of Pathology

    Article Title: Blood Vascular Abnormalities in Rasa1R780Q Knockin Mice

    doi: 10.1016/j.ajpath.2014.08.018

    Figure Lengend Snippet: MAPK activation in homozygous Rasa1 R780Q embryos. Tissue sections of E9.5 Rasa1 R780Q/R780Q and Rasa1 fl/fl embryos were stained with anti-CD31 (red) and anti–phospho-ERK (green) antibodies. Shown are select regions of embryos that include part of the dorsal aorta ( arrows ). Note the absence of pERK staining in most ECs in Rasa1 fl/fl embryos and the presence of pERK staining in most ECs in Rasa1 R780Q/R780Q embryos.

    Article Snippet: Fixed embryos were stained with a rat anti-CD31 antibody (BD Biosciences, San Jose, CA), followed by a secondary goat anti-rat Ig Alexa Fluor 488 antibody (Invitrogen, Carlsbad, CA).

    Techniques: Activation Assay, Staining

    Assessment of tumor malignancy based on CD31, MMP13 and TGF-β1 expression. (A) H E staining of sliced tissue samples from Non-Rec and Rec tumors. Areas containing angiogenic features are demarcated by the red lines and indicated with yellow arrows. The yellow arrow-heads indicated the elongated fibroblast. An enlarged view of the angiogenic area is shown to the right of each sample. (B) Immunofluorescent staining of Non-Rec and Rec tumor tissue samples for CD31 and MMP13. Colocalized staining of CD31 and MMP13 is indicated by the white arrows. (C) The bar graph demonstrates the difference in microvascular density (MVD) between the groups. (D) The ELISA assay showed that TGF-β1 was expressed in a recurrence-status-dependent manner, because the level of released TGF-β1 was lower in the culture medium from the Non-Rec group than from the Rec group. Scale bar = 100 μm in the H E images; Scale bar = 200 μm in the Immunofluorescent images. *, p

    Journal: Cancer Cell International

    Article Title: Validation of the effects of TGF-?1 on tumor recurrence and prognosis through tumor retrieval and cell mechanical properties

    doi: 10.1186/1475-2867-14-20

    Figure Lengend Snippet: Assessment of tumor malignancy based on CD31, MMP13 and TGF-β1 expression. (A) H E staining of sliced tissue samples from Non-Rec and Rec tumors. Areas containing angiogenic features are demarcated by the red lines and indicated with yellow arrows. The yellow arrow-heads indicated the elongated fibroblast. An enlarged view of the angiogenic area is shown to the right of each sample. (B) Immunofluorescent staining of Non-Rec and Rec tumor tissue samples for CD31 and MMP13. Colocalized staining of CD31 and MMP13 is indicated by the white arrows. (C) The bar graph demonstrates the difference in microvascular density (MVD) between the groups. (D) The ELISA assay showed that TGF-β1 was expressed in a recurrence-status-dependent manner, because the level of released TGF-β1 was lower in the culture medium from the Non-Rec group than from the Rec group. Scale bar = 100 μm in the H E images; Scale bar = 200 μm in the Immunofluorescent images. *, p

    Article Snippet: To evaluate tumor angiogenesis and invasiveness, tumor sections were stained with rat anti-mouse CD31 (1:100; Pharmingen, Heidelberg, Germany) and rabbit anti-mouse MMP-13 (1:500; Santa Cruz Biotechnology, Heidelberg, Germany) Subsequently, the sections were washed with PBS and incubated with an Alexa568-conjugated goat anti-rat secondary antibody (for CD31; 1:400; Dianova, Hamburg, Germany), and an Alexa488-conjugated goat anti-rabbit secondary antibody (for MMP13; 1:400; Molecular Probes, Göttingen, Germany) for 2 hr at room temperature.

    Techniques: Expressing, Staining, Enzyme-linked Immunosorbent Assay

    The confocal imaging of tumor, spleen, and lymph tissues from the MDA-MB-231 breast cancer model after immuno-fluorescent staining. Sample groups: DAPI, the nucleus stained by DAPI; PD-L1, the PD-L1 expression stained by avelumab as the primary antibody; CD31, the expression of vascular endothelium biomarker CD31.

    Journal: American Journal of Translational Research

    Article Title: In vivo characterization of PD-L1 expression in breast cancer by immuno-PET with 89Zr-labeled avelumab

    doi:

    Figure Lengend Snippet: The confocal imaging of tumor, spleen, and lymph tissues from the MDA-MB-231 breast cancer model after immuno-fluorescent staining. Sample groups: DAPI, the nucleus stained by DAPI; PD-L1, the PD-L1 expression stained by avelumab as the primary antibody; CD31, the expression of vascular endothelium biomarker CD31.

    Article Snippet: Pharmingen rat anti-mouse CD31 antibody was purchased from BD Bioscience, Inc. (San Diego, CA).

    Techniques: Imaging, Multiple Displacement Amplification, Staining, Expressing, Biomarker Assay

    Hemilimbal deficiency: a model for injury-induced corneal NV. Diagrams depict limbal injury (A; green) and limbal plus epithelial removal (D; purple). The nasal limbi of wild-type mouse corneas were removed and photographed at day 7 after surgery (B, C). The nasal limbus and the epithelium of WT mouse corneas were removed and the corneas photographed at day 7 after surgery (E, F). Vascularized vessels were immunostained with anti-type IV collagen (G), anti-CD31 antibodies (H), and double staining (I).

    Journal: Transactions of the American Ophthalmological Society

    Article Title: CORNEAL ANGIOGENIC PRIVILEGE: ANGIOGENIC AND ANTIANGIOGENIC FACTORS IN CORNEAL AVASCULARITY, VASCULOGENESIS, AND WOUND HEALING (AN AMERICAN OPHTHALMOLOGICAL SOCIETY THESIS)

    doi:

    Figure Lengend Snippet: Hemilimbal deficiency: a model for injury-induced corneal NV. Diagrams depict limbal injury (A; green) and limbal plus epithelial removal (D; purple). The nasal limbi of wild-type mouse corneas were removed and photographed at day 7 after surgery (B, C). The nasal limbus and the epithelium of WT mouse corneas were removed and the corneas photographed at day 7 after surgery (E, F). Vascularized vessels were immunostained with anti-type IV collagen (G), anti-CD31 antibodies (H), and double staining (I).

    Article Snippet: A blocking solution (1% BSA in PBS) was applied for 30 mintues at room temperature and followed by incubation with primary antibodies, rat anti-mouse CD31 monoclonal antibody (PharMingen, San Diego, California), and goat anti-mouse VEGF monoclonal antibody (Calbiochem) of 1:100 dilution for 60 minutes.

    Techniques: Double Staining

    Colocalization of MMP-2 and MMP-14 in vascular endothelial cells. Vascularized corneal vessels were immunostained with anti-MT1-MMP(A) and CD31 antibodies (B) and double stained with both (C). They were also immunostained with anti–type IV collagen (D) and anti-MMP-2 antibodies (E) and double stained with both (F).

    Journal: Transactions of the American Ophthalmological Society

    Article Title: CORNEAL ANGIOGENIC PRIVILEGE: ANGIOGENIC AND ANTIANGIOGENIC FACTORS IN CORNEAL AVASCULARITY, VASCULOGENESIS, AND WOUND HEALING (AN AMERICAN OPHTHALMOLOGICAL SOCIETY THESIS)

    doi:

    Figure Lengend Snippet: Colocalization of MMP-2 and MMP-14 in vascular endothelial cells. Vascularized corneal vessels were immunostained with anti-MT1-MMP(A) and CD31 antibodies (B) and double stained with both (C). They were also immunostained with anti–type IV collagen (D) and anti-MMP-2 antibodies (E) and double stained with both (F).

    Article Snippet: A blocking solution (1% BSA in PBS) was applied for 30 mintues at room temperature and followed by incubation with primary antibodies, rat anti-mouse CD31 monoclonal antibody (PharMingen, San Diego, California), and goat anti-mouse VEGF monoclonal antibody (Calbiochem) of 1:100 dilution for 60 minutes.

    Techniques: Staining

    Hemilimbal injury (HLD-) of corneas in wild-type, MMP-2−/−, and MMP12−/− mice. Hemilimbal injury was applied to mouse cornea in MMP-2−/− (A, B, and C), MMP12−/− (D, E, F), and wild-type mice (G, H, I) at day 7 after wounding. Corneal vessels were visualized with type IV collagen (J) and CD31 (K) and then the images were merged (L).

    Journal: Transactions of the American Ophthalmological Society

    Article Title: CORNEAL ANGIOGENIC PRIVILEGE: ANGIOGENIC AND ANTIANGIOGENIC FACTORS IN CORNEAL AVASCULARITY, VASCULOGENESIS, AND WOUND HEALING (AN AMERICAN OPHTHALMOLOGICAL SOCIETY THESIS)

    doi:

    Figure Lengend Snippet: Hemilimbal injury (HLD-) of corneas in wild-type, MMP-2−/−, and MMP12−/− mice. Hemilimbal injury was applied to mouse cornea in MMP-2−/− (A, B, and C), MMP12−/− (D, E, F), and wild-type mice (G, H, I) at day 7 after wounding. Corneal vessels were visualized with type IV collagen (J) and CD31 (K) and then the images were merged (L).

    Article Snippet: A blocking solution (1% BSA in PBS) was applied for 30 mintues at room temperature and followed by incubation with primary antibodies, rat anti-mouse CD31 monoclonal antibody (PharMingen, San Diego, California), and goat anti-mouse VEGF monoclonal antibody (Calbiochem) of 1:100 dilution for 60 minutes.

    Techniques: Mouse Assay

    Distribution of MT1-MMP, VEGF, and CD31 in mouse corneas 4 days following bFGF pellet implantation. FGF-2 pellets (50 ng) were implanted into mouse corneas. The corneas were harvested at 4 days postoperatively. The central avascular zone adjacent to the pellets and the peripheral neovascularization zone were examined. Individual sections were immunostained with MT1-MMP (A, B), VEGF (C, D), and CD31 (E, F), a vascular endothelial cell marker.

    Journal: Transactions of the American Ophthalmological Society

    Article Title: CORNEAL ANGIOGENIC PRIVILEGE: ANGIOGENIC AND ANTIANGIOGENIC FACTORS IN CORNEAL AVASCULARITY, VASCULOGENESIS, AND WOUND HEALING (AN AMERICAN OPHTHALMOLOGICAL SOCIETY THESIS)

    doi:

    Figure Lengend Snippet: Distribution of MT1-MMP, VEGF, and CD31 in mouse corneas 4 days following bFGF pellet implantation. FGF-2 pellets (50 ng) were implanted into mouse corneas. The corneas were harvested at 4 days postoperatively. The central avascular zone adjacent to the pellets and the peripheral neovascularization zone were examined. Individual sections were immunostained with MT1-MMP (A, B), VEGF (C, D), and CD31 (E, F), a vascular endothelial cell marker.

    Article Snippet: A blocking solution (1% BSA in PBS) was applied for 30 mintues at room temperature and followed by incubation with primary antibodies, rat anti-mouse CD31 monoclonal antibody (PharMingen, San Diego, California), and goat anti-mouse VEGF monoclonal antibody (Calbiochem) of 1:100 dilution for 60 minutes.

    Techniques: Marker

    Distribution of MT1-MMP, VEGF, and CD31 in mouse corneas 14 days after bFGF pellet implantation. FGF-2 pellets (50 ng) were implanted into mouse corneas. The corneas were harvested at 14 days postoperatively. The central avascular zone adjacent to the pellets and the peripheral neovascularization zone were examined. Individual sections were immunostained with MT1-MMP (A, B), VEGF (C, D), and CD31 (E, F), a vascular endothelial cell marker.

    Journal: Transactions of the American Ophthalmological Society

    Article Title: CORNEAL ANGIOGENIC PRIVILEGE: ANGIOGENIC AND ANTIANGIOGENIC FACTORS IN CORNEAL AVASCULARITY, VASCULOGENESIS, AND WOUND HEALING (AN AMERICAN OPHTHALMOLOGICAL SOCIETY THESIS)

    doi:

    Figure Lengend Snippet: Distribution of MT1-MMP, VEGF, and CD31 in mouse corneas 14 days after bFGF pellet implantation. FGF-2 pellets (50 ng) were implanted into mouse corneas. The corneas were harvested at 14 days postoperatively. The central avascular zone adjacent to the pellets and the peripheral neovascularization zone were examined. Individual sections were immunostained with MT1-MMP (A, B), VEGF (C, D), and CD31 (E, F), a vascular endothelial cell marker.

    Article Snippet: A blocking solution (1% BSA in PBS) was applied for 30 mintues at room temperature and followed by incubation with primary antibodies, rat anti-mouse CD31 monoclonal antibody (PharMingen, San Diego, California), and goat anti-mouse VEGF monoclonal antibody (Calbiochem) of 1:100 dilution for 60 minutes.

    Techniques: Marker

    Tumor vessel perfusion. The vascular function is increased in tumors of RT pos SG ko versus RT pos SG wt mice. 15 week RT pos SG wt (a) and RT pos SG ko (b) mice were perfused with FITC-lectin, which was introduced to the circulation prior to sacrifice of the animals and excision of the pancreas. Tumor sections were stained for the endothelial marker CD31 (red), the localization of which was compared to lectin (green) that had immobilized on the luminal side of endothelial cells in perfused vessels. The ratio between immobilized lectin and CD31 staining was calculated to determine the fraction of perfused, and thus functional vessels (c). Each data point in represents an individual animal. Statistical analysis was performed using a two-tailed Mann-Whitney test. Error bars represent mean ± SEM.

    Journal: PLoS ONE

    Article Title: Loss of Serglycin Promotes Primary Tumor Growth and Vessel Functionality in the RIP1-Tag2 Mouse Model for Spontaneous Insulinoma Formation

    doi: 10.1371/journal.pone.0126688

    Figure Lengend Snippet: Tumor vessel perfusion. The vascular function is increased in tumors of RT pos SG ko versus RT pos SG wt mice. 15 week RT pos SG wt (a) and RT pos SG ko (b) mice were perfused with FITC-lectin, which was introduced to the circulation prior to sacrifice of the animals and excision of the pancreas. Tumor sections were stained for the endothelial marker CD31 (red), the localization of which was compared to lectin (green) that had immobilized on the luminal side of endothelial cells in perfused vessels. The ratio between immobilized lectin and CD31 staining was calculated to determine the fraction of perfused, and thus functional vessels (c). Each data point in represents an individual animal. Statistical analysis was performed using a two-tailed Mann-Whitney test. Error bars represent mean ± SEM.

    Article Snippet: Immunogen: synthetic peptide corresponding to Human active + pro Caspase 3 residues 150–250 conjugated to KLH (mouse cross-reactivity)); Rat monoclonal anti-CD31, final dilution 1:1000 (557355, BD Pharmingen.

    Techniques: Mouse Assay, Staining, Marker, Functional Assay, Two Tailed Test, MANN-WHITNEY

    Representative examples of flow cytometry of CD42b–/CD31+ EMP in a rat exposed to smoke 2 months (B) 4 months (C) and 6 months (D) and a rat of sham exposure groups (A). The points represented in region Q4 of both examples are CD31+ (positive)

    Journal: Journal of Thoracic Disease

    Article Title: Circulating endothelial microparticles involved in lung function decline in a rat exposed in cigarette smoke maybe from apoptotic pulmonary capillary endothelial cells

    doi: 10.3978/j.issn.2072-1439.2014.06.26

    Figure Lengend Snippet: Representative examples of flow cytometry of CD42b–/CD31+ EMP in a rat exposed to smoke 2 months (B) 4 months (C) and 6 months (D) and a rat of sham exposure groups (A). The points represented in region Q4 of both examples are CD31+ (positive)

    Article Snippet: Terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) assays for apoptosis were performed referencing per manufacturer’s instructions (Roche, Indianapolis IN, USA), and subsequent staining with rat anti-CD31 monoclonal antibody (1:1,000, BD, USA) and Hoeschts dye (BisBenzimide, 1 mg/100 mL) were co-localized endothelial cells undergoing apoptosis by confocal microscopy.

    Techniques: Flow Cytometry, Cytometry

    Lung endothelial cell apoptosis after smoke exposure and sham exposure. Lung immunofluorescence micrographs (×200) stained with CD31 (green), TUNEL (red), Hoechst (blue), and merged CD31-TUNEL- Hoechst. The pink signals indicated by the arrows

    Journal: Journal of Thoracic Disease

    Article Title: Circulating endothelial microparticles involved in lung function decline in a rat exposed in cigarette smoke maybe from apoptotic pulmonary capillary endothelial cells

    doi: 10.3978/j.issn.2072-1439.2014.06.26

    Figure Lengend Snippet: Lung endothelial cell apoptosis after smoke exposure and sham exposure. Lung immunofluorescence micrographs (×200) stained with CD31 (green), TUNEL (red), Hoechst (blue), and merged CD31-TUNEL- Hoechst. The pink signals indicated by the arrows

    Article Snippet: Terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) assays for apoptosis were performed referencing per manufacturer’s instructions (Roche, Indianapolis IN, USA), and subsequent staining with rat anti-CD31 monoclonal antibody (1:1,000, BD, USA) and Hoeschts dye (BisBenzimide, 1 mg/100 mL) were co-localized endothelial cells undergoing apoptosis by confocal microscopy.

    Techniques: Immunofluorescence, Staining, TUNEL Assay

    Loss of Vav3 results in microvascular leakage in vivo. (A) Enhanced extravasation of red fluorescent microspheres in the trachea of Vav3 -deficient mice. Control and Vav3 −/− mice were injected (i.v.) with 23-nm red fluorescent microspheres (red) in combination with VEGF (3 μg per animal). Whole-mount preparations of trachea were immunostained with anti-CD31 (green). Bottom panels show magnification of boxed area in top panels (bars, 50 µm). (B) Quantification of microvascular leakage by determining area of microsphere extravasation versus total vessel area. Percentage of microsphere extravasation is set to 100% for trachea WT controls compared with Vav3 −/− (mean ± SEM; *, P

    Journal: The Journal of Cell Biology

    Article Title: Vav3-induced cytoskeletal dynamics contribute to heterotypic properties of endothelial barriers

    doi: 10.1083/jcb.201706041

    Figure Lengend Snippet: Loss of Vav3 results in microvascular leakage in vivo. (A) Enhanced extravasation of red fluorescent microspheres in the trachea of Vav3 -deficient mice. Control and Vav3 −/− mice were injected (i.v.) with 23-nm red fluorescent microspheres (red) in combination with VEGF (3 μg per animal). Whole-mount preparations of trachea were immunostained with anti-CD31 (green). Bottom panels show magnification of boxed area in top panels (bars, 50 µm). (B) Quantification of microvascular leakage by determining area of microsphere extravasation versus total vessel area. Percentage of microsphere extravasation is set to 100% for trachea WT controls compared with Vav3 −/− (mean ± SEM; *, P

    Article Snippet: Tracheas were dissected and fixed in 2% PFA over 2 h after a permeabilization and blocking step with 0.3% Triton X-100 and 3% donkey serum in PBS for 1 h. Tissue samples were incubated with rat anti–mouse CD31 (1:50; clone SZ31; Dianova) overnight and with Alexa Fluor secondary antibodies (1:400; Invitrogen), and DAPI (1:1,000) over 2 h the next day.

    Techniques: In Vivo, Mouse Assay, Injection

    CITED2 silencing attenuates MDA-MB-231, but not MDA-MB-468 tumor vascular area and diameter ( A and C ) Immunohistochemical analysis of CD31 protein expression (brown) in scramble and shCITED2-expressing tumors. Magnification: 100X. ( B and D ) Histograms representing the average overall number of blood vessels, vessel area and vessel diameter in each experimental group. (*** P

    Journal: Oncotarget

    Article Title: Down-regulation of CITED2 attenuates breast tumor growth, vessel formation and TGF-β-induced expression of VEGFA

    doi: 10.18632/oncotarget.14048

    Figure Lengend Snippet: CITED2 silencing attenuates MDA-MB-231, but not MDA-MB-468 tumor vascular area and diameter ( A and C ) Immunohistochemical analysis of CD31 protein expression (brown) in scramble and shCITED2-expressing tumors. Magnification: 100X. ( B and D ) Histograms representing the average overall number of blood vessels, vessel area and vessel diameter in each experimental group. (*** P

    Article Snippet: Sections were incubated with protein blocking solution (DAKO) for 20 minutes at room temperature followed by incubation at 4°C for 18 hours with rat anti-mouse CD31 antibody (1:30 dilution; DIA310, Dianova).

    Techniques: Multiple Displacement Amplification, Immunohistochemistry, Expressing

    Whole mount imaging of meningeal lymphatic vessels in mice. (a) Meningeal LVs were characterized by whole-mount immunofluorescence staining for CD31 (green) and Lyve-1 (red). Low magnification view of meningeal LVs. LVs run alongside the sagittal sinus (ss) and transverse sinus (ts). The boxed regions in panel A outline the areas that we analyzed in our stroke experiments to acquire lymphatic vessel index (LVI) at the ss. (b) Representative images showing Prox1 (red) and podoplanin (green) expression by meningeal LVs around the transverse sinus (TS). (c) A 3D rendering of meningeal LVs (red) in a Prox1-tdTomato transgenic mouse was generated using serial two-photon tomography. The signal from the hippocampus was digitally removed. (a), scale bar is 1000 µm; (b), scale bar is 20 µm.

    Journal: Journal of Cerebral Blood Flow & Metabolism

    Article Title: Impaired meningeal lymphatic vessel development worsens stroke outcome

    doi: 10.1177/0271678X18822921

    Figure Lengend Snippet: Whole mount imaging of meningeal lymphatic vessels in mice. (a) Meningeal LVs were characterized by whole-mount immunofluorescence staining for CD31 (green) and Lyve-1 (red). Low magnification view of meningeal LVs. LVs run alongside the sagittal sinus (ss) and transverse sinus (ts). The boxed regions in panel A outline the areas that we analyzed in our stroke experiments to acquire lymphatic vessel index (LVI) at the ss. (b) Representative images showing Prox1 (red) and podoplanin (green) expression by meningeal LVs around the transverse sinus (TS). (c) A 3D rendering of meningeal LVs (red) in a Prox1-tdTomato transgenic mouse was generated using serial two-photon tomography. The signal from the hippocampus was digitally removed. (a), scale bar is 1000 µm; (b), scale bar is 20 µm.

    Article Snippet: The following primary antibodies were used: rat anti-CD31 (1:1000; eBiosciences, San Diego, CA, USA), goat anti-Lyve-1 (1:1000; R & D Systems, Minneapolis, MN, USA), hamster anti-podoplanin (1:1000; Abcam, San Francisco, CA, USA), and rabbit anti-Prox1 (1:500; AngioBioCo, Del Mar, CA, USA).

    Techniques: Imaging, Mouse Assay, Immunofluorescence, Staining, Expressing, Transgenic Assay, Generated

    PLX4720 treatment stabilizes blood vessels in COLO205 and SK-MEL-28 xenograft models. ( A ) Representative images of immunofluorescence analysis with the CD31 endothelial marker (green) in COLO205 ( a – c ) and SK-MEL-28 ( d – f ) tumors. (Scale

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Targeting oncogenic serine/threonine-protein kinase BRAF in cancer cells inhibits angiogenesis and abrogates hypoxia

    doi: 10.1073/pnas.1105026109

    Figure Lengend Snippet: PLX4720 treatment stabilizes blood vessels in COLO205 and SK-MEL-28 xenograft models. ( A ) Representative images of immunofluorescence analysis with the CD31 endothelial marker (green) in COLO205 ( a – c ) and SK-MEL-28 ( d – f ) tumors. (Scale

    Article Snippet: The following primary antibodies were used for histological analysis: rabbit anti-von Willebrand Factor (vWF; A0082; DakoCytomation), mouse anti–HLA-ABC antigen (M0736; DakoCytomation), rabbit anti-Ki67 (RM-9106-F0; Thermo Scientific) rat anti-CD31 (Pecam-1; 550274BD; Pharmingen), anti-NG2 (chondroitin sulfate proteoglycan polyclonal; AB5320; Chemicon).

    Techniques: Immunofluorescence, Marker

    F8-VEGFC induces expansion of the lymphatic vasculature in chronic colitis. (A) Representative pictures of immunofluorescence labeling of the inflamed colon after F8-SIP (upper panels) or F8-VEGFC (lower panels) treatment for LYVE-1 (green), CD31 (red), and Hoechst (blue). Dotted white lines mark the separation between the mucosa (M) and the submucosa (SM). Scale bars: 100 μm. LYVE-1 + LVs were quantified in terms of number (B) and percentage of stained area (C) in 5–8 fields per colon per mouse ( n = 5 per group). CD31+LYVE-1- blood vessels were quantified in terms of number (D) and percentage of stained area (E) in 5–8 fields per colon per mouse ( n = 5 per group). All data are presented as mean ± SD. Statistical significance was determined by two-tailed Student’s t test. Asterisks indicate statistical significance with p

    Journal: ACS Pharmacology & Translational Science

    Article Title: Antibody-Mediated Delivery of VEGFC Ameliorates Experimental Chronic Colitis

    doi: 10.1021/acsptsci.9b00037

    Figure Lengend Snippet: F8-VEGFC induces expansion of the lymphatic vasculature in chronic colitis. (A) Representative pictures of immunofluorescence labeling of the inflamed colon after F8-SIP (upper panels) or F8-VEGFC (lower panels) treatment for LYVE-1 (green), CD31 (red), and Hoechst (blue). Dotted white lines mark the separation between the mucosa (M) and the submucosa (SM). Scale bars: 100 μm. LYVE-1 + LVs were quantified in terms of number (B) and percentage of stained area (C) in 5–8 fields per colon per mouse ( n = 5 per group). CD31+LYVE-1- blood vessels were quantified in terms of number (D) and percentage of stained area (E) in 5–8 fields per colon per mouse ( n = 5 per group). All data are presented as mean ± SD. Statistical significance was determined by two-tailed Student’s t test. Asterisks indicate statistical significance with p

    Article Snippet: After fixation in acetone and rehydration in 80% methanol, the sections were incubated with blocking solution (5% donkey serum, 1% bovine serum albumin (BSA), 0.1% Triton-X in PBS) for 1 h. EDA-FN staining was performed using a biotinylated F8 antibody in the small immunoprotein (SIP) format which was self-produced (400 μg/mL) and a rat anti-CD31 (Dianova DIA-310, 1:20) primary antibody in blocking solution at 4 °C overnight.

    Techniques: Immunofluorescence, Labeling, Staining, Two Tailed Test

    EDA-FN is increased in inflamed colon tissue. (A) EDA-FN expression levels were assessed in healthy ( n = 6), ulcerative colitis (UC, n = 24), and Crohn’s disease (CD, n = 19) endoscopic colonic biopsies, and absolute values for the probe 212464_s_at, recognizing EDA-FN, are shown. (B) Representative immunofluorescence pictures of naïve (CTRL, n = 3) and inflamed colons at day 24 post DSS administration (DSS, n = 3) stained for CD31 (red), EDA-FN (green), and Hoechst (blue). Scale bar: 100 μm. All data are presented as mean ± SD. Adjusted p value * p

    Journal: ACS Pharmacology & Translational Science

    Article Title: Antibody-Mediated Delivery of VEGFC Ameliorates Experimental Chronic Colitis

    doi: 10.1021/acsptsci.9b00037

    Figure Lengend Snippet: EDA-FN is increased in inflamed colon tissue. (A) EDA-FN expression levels were assessed in healthy ( n = 6), ulcerative colitis (UC, n = 24), and Crohn’s disease (CD, n = 19) endoscopic colonic biopsies, and absolute values for the probe 212464_s_at, recognizing EDA-FN, are shown. (B) Representative immunofluorescence pictures of naïve (CTRL, n = 3) and inflamed colons at day 24 post DSS administration (DSS, n = 3) stained for CD31 (red), EDA-FN (green), and Hoechst (blue). Scale bar: 100 μm. All data are presented as mean ± SD. Adjusted p value * p

    Article Snippet: After fixation in acetone and rehydration in 80% methanol, the sections were incubated with blocking solution (5% donkey serum, 1% bovine serum albumin (BSA), 0.1% Triton-X in PBS) for 1 h. EDA-FN staining was performed using a biotinylated F8 antibody in the small immunoprotein (SIP) format which was self-produced (400 μg/mL) and a rat anti-CD31 (Dianova DIA-310, 1:20) primary antibody in blocking solution at 4 °C overnight.

    Techniques: Expressing, Immunofluorescence, Staining

    A model for the CD31-induced barrier response. MHC triggering induces RhoA and Erk activation and EC contraction (1). Erk phosphorylation is modulated by CD31 signals, possibly via SHP-2 (2). MHC signals induce CD31 ITIM phosphorylation and SHP-2 recruitment. SHP-2 prevents the phosphorylation of b-catenin (5) and VE-cadherin (6), thus stabilizing the junctional complex. In addition, dephosphorylated b-catenin can transfer to the nucleus where it induces cMyc transcription. In parallel, SHP-2 induces AKT activation which in turn inhibits FoxO1 nuclear translocation, thus preventing inhibition of cMyc transcription. This leads to enhanced transcription of glycolysis enzymes and enhanced glycolysis required for actin remodeling and maintenance of junctional anchorage.

    Journal: Nature Communications

    Article Title: Preservation of microvascular barrier function requires CD31 receptor-induced metabolic reprogramming

    doi: 10.1038/s41467-020-17329-8

    Figure Lengend Snippet: A model for the CD31-induced barrier response. MHC triggering induces RhoA and Erk activation and EC contraction (1). Erk phosphorylation is modulated by CD31 signals, possibly via SHP-2 (2). MHC signals induce CD31 ITIM phosphorylation and SHP-2 recruitment. SHP-2 prevents the phosphorylation of b-catenin (5) and VE-cadherin (6), thus stabilizing the junctional complex. In addition, dephosphorylated b-catenin can transfer to the nucleus where it induces cMyc transcription. In parallel, SHP-2 induces AKT activation which in turn inhibits FoxO1 nuclear translocation, thus preventing inhibition of cMyc transcription. This leads to enhanced transcription of glycolysis enzymes and enhanced glycolysis required for actin remodeling and maintenance of junctional anchorage.

    Article Snippet: Mouse-anti-mouse H-2Ld/H-2Db (MHC-class-I) Antibody (Biolegend, 114502), Rat-anti-mouse IgG, (Biolegend, 553445), (Blocking) rat anti-mouse CD31 clone 390 (eBioscience, AB10060377), (Stimulate) rabbit anti-mouse CD31 (Abcam, ab2836), Goat–anti-rabbit Ig Antibody, (Agrisera, AS10 665), (cross-linkage) Mouse-anti-mouse ICAM-1 (Abcam, ab2213) were used to stimulate MHC molecules on the endothelium.

    Techniques: Activation Assay, Translocation Assay, Inhibition

    The metabolic response to endothelial barrier breach is src-phosphatase-dependent. Confluent EC monolayers were MHC-stimulated and treated with the selective SHP1/2 inhibitor (50 μM for 3 h) or vehicle as a control. a The bar graph shows the mean TEER values measured in three independent experiments. b Percentage of EC viability following treatment with SHP1/2 inhibitor determined by the trypan blue exclusion assay in three independent experiments. c – f ECAR measured in WT ( c , d ) or cd31 −/− EC ( e , f ) following MHC antibody-stimulation with or without pre-treatment with the SHP1/2 inhibitor (50μM for 45 min). Basal and maximal glycolysis and glycolytic reserve are shown in ( d , f ). g – j OCR measured in WT g – h or CD31-deficient EC I , j following MHC antibody-stimulation with or without pre-treatment with the SHP1/2 inhibitor. Basal and maximal respiration and spare capacity are shown in ( i , j ). ( N = 3 independent experiments). Data are shown as mean ± SEM; significant differences were determined using one-way Anova with Tuckey post-hoc test. a IsC vehicle vs MHC vehicle **** p

    Journal: Nature Communications

    Article Title: Preservation of microvascular barrier function requires CD31 receptor-induced metabolic reprogramming

    doi: 10.1038/s41467-020-17329-8

    Figure Lengend Snippet: The metabolic response to endothelial barrier breach is src-phosphatase-dependent. Confluent EC monolayers were MHC-stimulated and treated with the selective SHP1/2 inhibitor (50 μM for 3 h) or vehicle as a control. a The bar graph shows the mean TEER values measured in three independent experiments. b Percentage of EC viability following treatment with SHP1/2 inhibitor determined by the trypan blue exclusion assay in three independent experiments. c – f ECAR measured in WT ( c , d ) or cd31 −/− EC ( e , f ) following MHC antibody-stimulation with or without pre-treatment with the SHP1/2 inhibitor (50μM for 45 min). Basal and maximal glycolysis and glycolytic reserve are shown in ( d , f ). g – j OCR measured in WT g – h or CD31-deficient EC I , j following MHC antibody-stimulation with or without pre-treatment with the SHP1/2 inhibitor. Basal and maximal respiration and spare capacity are shown in ( i , j ). ( N = 3 independent experiments). Data are shown as mean ± SEM; significant differences were determined using one-way Anova with Tuckey post-hoc test. a IsC vehicle vs MHC vehicle **** p

    Article Snippet: Mouse-anti-mouse H-2Ld/H-2Db (MHC-class-I) Antibody (Biolegend, 114502), Rat-anti-mouse IgG, (Biolegend, 553445), (Blocking) rat anti-mouse CD31 clone 390 (eBioscience, AB10060377), (Stimulate) rabbit anti-mouse CD31 (Abcam, ab2836), Goat–anti-rabbit Ig Antibody, (Agrisera, AS10 665), (cross-linkage) Mouse-anti-mouse ICAM-1 (Abcam, ab2213) were used to stimulate MHC molecules on the endothelium.

    Techniques: Trypan Blue Exclusion Assay

    Akt and AMPK activation restore the glycolytic response in CD31-deficient EC in-vitro and in-vivo. cd31 −/− EC were treated with an Akt activator (500 nM, 3 h, a ) or Metformin (5 mM, 3 h, b ), prior to MHC-stimulation and glycolytic flux measurement N = 3 independent experiments. Error bars represent SD. One-way Anova with Tuckey post-hoc test. a Glu/Oligo injection MHC + Akt activator vs all * p = 0.04, Oligo/2DG injection MHC + Akt activator vs IsC * p = 0.048; b Glu/Oligo injection MHC + Metformin vs all ** p = 0.003, Oligo/2DG injection MHC + Metformin vs all * p = 0.05. c , d cd31 −/− EC were immunostained 2 h after MHC-stimulation by rabbit anti-mouse FoxO1 (green) and DAPI (blue). In some cultures, metformin was added. 500/coverslip were analyzed. d Percentage of cells displaying nuclear FoxO1 localization ( N = 3 independent experiments). Scale bar = 20 μm. e Quantitative analysis of TEER of cd31 −/− EC treated as indicated 3 h after stimulation. f CD31 −/− mice ( n = 5 mice, N = 2 independent experiments) received anti-MHC and secondary cross-linking antibody (0.67 μg and 0.33 μg/kg body weight, respectively) or an Isotype-matched control antibody (IsC) and secondary cross-linking antibody (3.35 μg and 1.7 μg/kg body weight, respectively) i.v. Some mice received either an Akt activator (i.p., 7 mg/kg) or Metformin (i.p., 125 mg/kg). After 6 h, Evans blue dye (2 mg/kg) was administered by i.v. and organ dye ( n = 3 mice; N = 2 independent experiments). Error bars represent SD. One-way Anova with Tuckey post-hoc test. d IsC vs MHC **** p

    Journal: Nature Communications

    Article Title: Preservation of microvascular barrier function requires CD31 receptor-induced metabolic reprogramming

    doi: 10.1038/s41467-020-17329-8

    Figure Lengend Snippet: Akt and AMPK activation restore the glycolytic response in CD31-deficient EC in-vitro and in-vivo. cd31 −/− EC were treated with an Akt activator (500 nM, 3 h, a ) or Metformin (5 mM, 3 h, b ), prior to MHC-stimulation and glycolytic flux measurement N = 3 independent experiments. Error bars represent SD. One-way Anova with Tuckey post-hoc test. a Glu/Oligo injection MHC + Akt activator vs all * p = 0.04, Oligo/2DG injection MHC + Akt activator vs IsC * p = 0.048; b Glu/Oligo injection MHC + Metformin vs all ** p = 0.003, Oligo/2DG injection MHC + Metformin vs all * p = 0.05. c , d cd31 −/− EC were immunostained 2 h after MHC-stimulation by rabbit anti-mouse FoxO1 (green) and DAPI (blue). In some cultures, metformin was added. 500/coverslip were analyzed. d Percentage of cells displaying nuclear FoxO1 localization ( N = 3 independent experiments). Scale bar = 20 μm. e Quantitative analysis of TEER of cd31 −/− EC treated as indicated 3 h after stimulation. f CD31 −/− mice ( n = 5 mice, N = 2 independent experiments) received anti-MHC and secondary cross-linking antibody (0.67 μg and 0.33 μg/kg body weight, respectively) or an Isotype-matched control antibody (IsC) and secondary cross-linking antibody (3.35 μg and 1.7 μg/kg body weight, respectively) i.v. Some mice received either an Akt activator (i.p., 7 mg/kg) or Metformin (i.p., 125 mg/kg). After 6 h, Evans blue dye (2 mg/kg) was administered by i.v. and organ dye ( n = 3 mice; N = 2 independent experiments). Error bars represent SD. One-way Anova with Tuckey post-hoc test. d IsC vs MHC **** p

    Article Snippet: Mouse-anti-mouse H-2Ld/H-2Db (MHC-class-I) Antibody (Biolegend, 114502), Rat-anti-mouse IgG, (Biolegend, 553445), (Blocking) rat anti-mouse CD31 clone 390 (eBioscience, AB10060377), (Stimulate) rabbit anti-mouse CD31 (Abcam, ab2836), Goat–anti-rabbit Ig Antibody, (Agrisera, AS10 665), (cross-linkage) Mouse-anti-mouse ICAM-1 (Abcam, ab2213) were used to stimulate MHC molecules on the endothelium.

    Techniques: Activation Assay, In Vitro, In Vivo, Injection, Mouse Assay

    CD31 signals induce transcription of glycolytic enzymes via inhibition of FoxO1 activity. WT and cd31 −/ − EC were stimulated by MHC antibody-ligation or treated with an isotype-matched control and secondary antibody (4 h). a , b Expression of the indicated enzymes was analyzed by immunoblotting and quantified by densitometry. N = 3 independent experiments. c Expression of aldolase A mRNA in WT and CD31 deficient EC was determined by RT-PCR 2 h after stimulation. N = 3 independent experiments. Data are mean ± SD. One-way Anova with Tuckey post-hoc test. b PFKFB3 WT IsC vs WT MHC **** p

    Journal: Nature Communications

    Article Title: Preservation of microvascular barrier function requires CD31 receptor-induced metabolic reprogramming

    doi: 10.1038/s41467-020-17329-8

    Figure Lengend Snippet: CD31 signals induce transcription of glycolytic enzymes via inhibition of FoxO1 activity. WT and cd31 −/ − EC were stimulated by MHC antibody-ligation or treated with an isotype-matched control and secondary antibody (4 h). a , b Expression of the indicated enzymes was analyzed by immunoblotting and quantified by densitometry. N = 3 independent experiments. c Expression of aldolase A mRNA in WT and CD31 deficient EC was determined by RT-PCR 2 h after stimulation. N = 3 independent experiments. Data are mean ± SD. One-way Anova with Tuckey post-hoc test. b PFKFB3 WT IsC vs WT MHC **** p

    Article Snippet: Mouse-anti-mouse H-2Ld/H-2Db (MHC-class-I) Antibody (Biolegend, 114502), Rat-anti-mouse IgG, (Biolegend, 553445), (Blocking) rat anti-mouse CD31 clone 390 (eBioscience, AB10060377), (Stimulate) rabbit anti-mouse CD31 (Abcam, ab2836), Goat–anti-rabbit Ig Antibody, (Agrisera, AS10 665), (cross-linkage) Mouse-anti-mouse ICAM-1 (Abcam, ab2213) were used to stimulate MHC molecules on the endothelium.

    Techniques: Inhibition, Activity Assay, Ligation, Expressing, Reverse Transcription Polymerase Chain Reaction

    CD31 signaling promotes β-catenin nuclear translocation and upregulates cMyc expression. WT and cd31 −/− EC monolayers were stimulated by MHC antibody-ligation or treated with an Isotype-matched control and secondary antibodies (2 h). Some cd31 −/− ECs were also treated with an Akt activator (500 nM, 3 h) prior to antibody stimulation. Vehicle was added in the untreated cultures (IsC and MHC ligation). a β-catenin and cMyc expression were determined by immunofluorescent antibody staining and wide-field microscopy. The nucleus was stained with DAPI. The mean fluorescence intensity of cMyc and β-catenin measured in 500 cells in three independent experiments is shown in ( b , c ), respectively. Scale bar = 40 μm. Data are mean ± SD. one-way Anova with Tuckey post-hoc test. b WT IsC vs WT MHC; c WT IsC vs WT MHC *** p = 0.0008, cd31 − / − IsC + C991 vs cd31 − / − MHC + C991 *** p = 0.0002. d , e : cMyc ( d ) and aldolase ( e ) gene transcription by WT (upper panels) and cd31 −/− (lower panels) EC at the indicated time points. n = 3 biologically independent samples, N = 2 independent experiments. Error bars represent SD. One-way Anova with Tuckey post-hoc test or T -test ( d , e ). d WT IsC 30′ vs WT MHC 30′ ** p = 0.003, WT IsC 120′ vs WT MHC 120′ **** p

    Journal: Nature Communications

    Article Title: Preservation of microvascular barrier function requires CD31 receptor-induced metabolic reprogramming

    doi: 10.1038/s41467-020-17329-8

    Figure Lengend Snippet: CD31 signaling promotes β-catenin nuclear translocation and upregulates cMyc expression. WT and cd31 −/− EC monolayers were stimulated by MHC antibody-ligation or treated with an Isotype-matched control and secondary antibodies (2 h). Some cd31 −/− ECs were also treated with an Akt activator (500 nM, 3 h) prior to antibody stimulation. Vehicle was added in the untreated cultures (IsC and MHC ligation). a β-catenin and cMyc expression were determined by immunofluorescent antibody staining and wide-field microscopy. The nucleus was stained with DAPI. The mean fluorescence intensity of cMyc and β-catenin measured in 500 cells in three independent experiments is shown in ( b , c ), respectively. Scale bar = 40 μm. Data are mean ± SD. one-way Anova with Tuckey post-hoc test. b WT IsC vs WT MHC; c WT IsC vs WT MHC *** p = 0.0008, cd31 − / − IsC + C991 vs cd31 − / − MHC + C991 *** p = 0.0002. d , e : cMyc ( d ) and aldolase ( e ) gene transcription by WT (upper panels) and cd31 −/− (lower panels) EC at the indicated time points. n = 3 biologically independent samples, N = 2 independent experiments. Error bars represent SD. One-way Anova with Tuckey post-hoc test or T -test ( d , e ). d WT IsC 30′ vs WT MHC 30′ ** p = 0.003, WT IsC 120′ vs WT MHC 120′ **** p

    Article Snippet: Mouse-anti-mouse H-2Ld/H-2Db (MHC-class-I) Antibody (Biolegend, 114502), Rat-anti-mouse IgG, (Biolegend, 553445), (Blocking) rat anti-mouse CD31 clone 390 (eBioscience, AB10060377), (Stimulate) rabbit anti-mouse CD31 (Abcam, ab2836), Goat–anti-rabbit Ig Antibody, (Agrisera, AS10 665), (cross-linkage) Mouse-anti-mouse ICAM-1 (Abcam, ab2213) were used to stimulate MHC molecules on the endothelium.

    Techniques: Translocation Assay, Expressing, Ligation, Staining, Microscopy, Fluorescence

    CD31 interactions promote the recovery of endothelial integrity following endothelial contraction induced by MHC molecule triggering. a – d Following MHC or ICAM-1 and/or CD31 antibody-mediated co-ligation for 30 min, EC were fixed and stained with rhodamine-phalloidin. Images taken on EC monolayers seeded at identical density are shown in ( a , b ). The average F-actin intensity per cell of three independent experiments is shown in ( c , d ). Scale bar, 20 μm. ( n = 3 biologically independent samples, N = 3 independent experiments, data are mean ± SD). One-way Anova with Tuckey post-hoc test. MHC vs all **** p

    Journal: Nature Communications

    Article Title: Preservation of microvascular barrier function requires CD31 receptor-induced metabolic reprogramming

    doi: 10.1038/s41467-020-17329-8

    Figure Lengend Snippet: CD31 interactions promote the recovery of endothelial integrity following endothelial contraction induced by MHC molecule triggering. a – d Following MHC or ICAM-1 and/or CD31 antibody-mediated co-ligation for 30 min, EC were fixed and stained with rhodamine-phalloidin. Images taken on EC monolayers seeded at identical density are shown in ( a , b ). The average F-actin intensity per cell of three independent experiments is shown in ( c , d ). Scale bar, 20 μm. ( n = 3 biologically independent samples, N = 3 independent experiments, data are mean ± SD). One-way Anova with Tuckey post-hoc test. MHC vs all **** p

    Article Snippet: Mouse-anti-mouse H-2Ld/H-2Db (MHC-class-I) Antibody (Biolegend, 114502), Rat-anti-mouse IgG, (Biolegend, 553445), (Blocking) rat anti-mouse CD31 clone 390 (eBioscience, AB10060377), (Stimulate) rabbit anti-mouse CD31 (Abcam, ab2836), Goat–anti-rabbit Ig Antibody, (Agrisera, AS10 665), (cross-linkage) Mouse-anti-mouse ICAM-1 (Abcam, ab2213) were used to stimulate MHC molecules on the endothelium.

    Techniques: Ligation, Staining

    CD31 signals prevent VE-cadherin and β-catenin phosphorylation in response to MHC stimulation via ITIM 686-thyrosine phosphorylation. a phosphorylation of β-catenin (Y654) by MHC- or ICAM-1-stimulated (30 min) WT or cd31 −/− EC was analyzed by western blotting. The bar graph shows relative protein expression ± SEM. ( N = 3 independent experiments). One-way Anova with Tuckey post-hoc test. cd31 −/− MHC vs all *** p = 0.0002. b – e phosphorylation of β-catenin (Y654, b ) or VE-cadherin (Y685, d ) by MHC-stimulated (30 min) WT or CD31 − / − EC was analyzed by widefield fluorescence microscopy. The bar graphs c , e show the mean fluorescence intensity/per cell of the indicated marker measured in three independent experiments by ImageJ software. Scale bar, 20 μm. Magnification ×20. Data are mean ± SD. One-way Anova with Tuckey post-hoc test. c cd31 − / − MHC vs all **** p

    Journal: Nature Communications

    Article Title: Preservation of microvascular barrier function requires CD31 receptor-induced metabolic reprogramming

    doi: 10.1038/s41467-020-17329-8

    Figure Lengend Snippet: CD31 signals prevent VE-cadherin and β-catenin phosphorylation in response to MHC stimulation via ITIM 686-thyrosine phosphorylation. a phosphorylation of β-catenin (Y654) by MHC- or ICAM-1-stimulated (30 min) WT or cd31 −/− EC was analyzed by western blotting. The bar graph shows relative protein expression ± SEM. ( N = 3 independent experiments). One-way Anova with Tuckey post-hoc test. cd31 −/− MHC vs all *** p = 0.0002. b – e phosphorylation of β-catenin (Y654, b ) or VE-cadherin (Y685, d ) by MHC-stimulated (30 min) WT or CD31 − / − EC was analyzed by widefield fluorescence microscopy. The bar graphs c , e show the mean fluorescence intensity/per cell of the indicated marker measured in three independent experiments by ImageJ software. Scale bar, 20 μm. Magnification ×20. Data are mean ± SD. One-way Anova with Tuckey post-hoc test. c cd31 − / − MHC vs all **** p

    Article Snippet: Mouse-anti-mouse H-2Ld/H-2Db (MHC-class-I) Antibody (Biolegend, 114502), Rat-anti-mouse IgG, (Biolegend, 553445), (Blocking) rat anti-mouse CD31 clone 390 (eBioscience, AB10060377), (Stimulate) rabbit anti-mouse CD31 (Abcam, ab2836), Goat–anti-rabbit Ig Antibody, (Agrisera, AS10 665), (cross-linkage) Mouse-anti-mouse ICAM-1 (Abcam, ab2213) were used to stimulate MHC molecules on the endothelium.

    Techniques: Western Blot, Expressing, Fluorescence, Microscopy, Marker, Software

    CD31-deficient EC increase mitochondrial respiration upon MHC stimulation. Oxygen consumption rate (OCR), basal, maximal respiration, and spare capacity of unstimulated WT and CD31-deficient EC is shown in ( a , b ), respectively ( N = 3 independent experiments). The error bars represent SD. One-way Anova with Tuckey post-hoc test. Basal respiration WT vs cd31 −/− **** p

    Journal: Nature Communications

    Article Title: Preservation of microvascular barrier function requires CD31 receptor-induced metabolic reprogramming

    doi: 10.1038/s41467-020-17329-8

    Figure Lengend Snippet: CD31-deficient EC increase mitochondrial respiration upon MHC stimulation. Oxygen consumption rate (OCR), basal, maximal respiration, and spare capacity of unstimulated WT and CD31-deficient EC is shown in ( a , b ), respectively ( N = 3 independent experiments). The error bars represent SD. One-way Anova with Tuckey post-hoc test. Basal respiration WT vs cd31 −/− **** p

    Article Snippet: Mouse-anti-mouse H-2Ld/H-2Db (MHC-class-I) Antibody (Biolegend, 114502), Rat-anti-mouse IgG, (Biolegend, 553445), (Blocking) rat anti-mouse CD31 clone 390 (eBioscience, AB10060377), (Stimulate) rabbit anti-mouse CD31 (Abcam, ab2836), Goat–anti-rabbit Ig Antibody, (Agrisera, AS10 665), (cross-linkage) Mouse-anti-mouse ICAM-1 (Abcam, ab2213) were used to stimulate MHC molecules on the endothelium.

    Techniques:

    CD31 regulates EBF via its effects on EC metabolism. a EC were pre-treated with the glucose analog 2-DG (5 mM, 2 h) then MHC-1- or ICAM–stimulated for 4 h prior to TEER measurements. The bar graph shows the mean of measurement collected in three separate experiments of identical design ± SD. One-way Anova with Tuckey post-hoc test. 2DG MHC vs all *** p = 0.0003. b Glut-1 expression by EC following MHC stimulation (2 h) was evaluated by qRT-PCR. Data are mean ± SD. Two-tailed Student’s T test. MHC vs IsC * p = 0.0412. c , d Representative histograms of antibody-stimulated WT or CD31-deficient EC incubated with 6-NBDG for 2 h prior to analysis. 2-DG was used as a negative control. The graph shows the mean MFI measured in three independent experiments ± SD. One-way Anova with Tuckey post-hoc test. WT IsC vs WT MHC ** p = 0.003, cd31 −/− IsC vs cd31 −/− MHC ns = 0.058. e ATP levels were measured in WT and CD31-deficient EC 4 h after MHC or insulin (1.8 µM) stimulation. Data are mean ± SD. Two-tailed Student’s T test. WT MHC vs cd31 −/− MHC *** p = 0.0002, WT insulin vs cd31 −/− insulin ** p = 0.0062. Extracellular acidification rate (ECAR) of unstimulated and antibody-stimulated WT and CD31-deficient EC is shown in ( f – h ). The basal and maximal glycolysis and the glycolytic reserve are shown in ( i – k ). Wells were first injected with anti MHC antibodies at the indicated time point. Isotype-matched and secondary antibodies were used as controls. Further injections followed at the time point indicated (arrows) introducing the indicated compounds into the wells. N = 3 independent experiments. The error bars represent SD. One-way Anova with Tuckey post-hoc test. f WT time 81 vs cd31 −/− time 81 ** p = 0.0023, WT time 94 vs cd31 −/− time 94 *** p = 0.0017; h IsC Glu/Oligo injection vs MHC Glu/Oligo ** p = 0.012, IsC Oligo/2DG vs MHC Oligo/2DG * p = 0.027; i WT MHC vs WT IsC **** p

    Journal: Nature Communications

    Article Title: Preservation of microvascular barrier function requires CD31 receptor-induced metabolic reprogramming

    doi: 10.1038/s41467-020-17329-8

    Figure Lengend Snippet: CD31 regulates EBF via its effects on EC metabolism. a EC were pre-treated with the glucose analog 2-DG (5 mM, 2 h) then MHC-1- or ICAM–stimulated for 4 h prior to TEER measurements. The bar graph shows the mean of measurement collected in three separate experiments of identical design ± SD. One-way Anova with Tuckey post-hoc test. 2DG MHC vs all *** p = 0.0003. b Glut-1 expression by EC following MHC stimulation (2 h) was evaluated by qRT-PCR. Data are mean ± SD. Two-tailed Student’s T test. MHC vs IsC * p = 0.0412. c , d Representative histograms of antibody-stimulated WT or CD31-deficient EC incubated with 6-NBDG for 2 h prior to analysis. 2-DG was used as a negative control. The graph shows the mean MFI measured in three independent experiments ± SD. One-way Anova with Tuckey post-hoc test. WT IsC vs WT MHC ** p = 0.003, cd31 −/− IsC vs cd31 −/− MHC ns = 0.058. e ATP levels were measured in WT and CD31-deficient EC 4 h after MHC or insulin (1.8 µM) stimulation. Data are mean ± SD. Two-tailed Student’s T test. WT MHC vs cd31 −/− MHC *** p = 0.0002, WT insulin vs cd31 −/− insulin ** p = 0.0062. Extracellular acidification rate (ECAR) of unstimulated and antibody-stimulated WT and CD31-deficient EC is shown in ( f – h ). The basal and maximal glycolysis and the glycolytic reserve are shown in ( i – k ). Wells were first injected with anti MHC antibodies at the indicated time point. Isotype-matched and secondary antibodies were used as controls. Further injections followed at the time point indicated (arrows) introducing the indicated compounds into the wells. N = 3 independent experiments. The error bars represent SD. One-way Anova with Tuckey post-hoc test. f WT time 81 vs cd31 −/− time 81 ** p = 0.0023, WT time 94 vs cd31 −/− time 94 *** p = 0.0017; h IsC Glu/Oligo injection vs MHC Glu/Oligo ** p = 0.012, IsC Oligo/2DG vs MHC Oligo/2DG * p = 0.027; i WT MHC vs WT IsC **** p

    Article Snippet: Mouse-anti-mouse H-2Ld/H-2Db (MHC-class-I) Antibody (Biolegend, 114502), Rat-anti-mouse IgG, (Biolegend, 553445), (Blocking) rat anti-mouse CD31 clone 390 (eBioscience, AB10060377), (Stimulate) rabbit anti-mouse CD31 (Abcam, ab2836), Goat–anti-rabbit Ig Antibody, (Agrisera, AS10 665), (cross-linkage) Mouse-anti-mouse ICAM-1 (Abcam, ab2213) were used to stimulate MHC molecules on the endothelium.

    Techniques: Expressing, Quantitative RT-PCR, Two Tailed Test, Incubation, Negative Control, Injection

    Enhanced rejection of CD31-deficient skin grafts. WT female mice received either a WT or CD31-KO male-derived skin graft. As a control, female WT and CD31-KO female-derived skin was used. ( A ) Graft survival ( n = 6). **** P

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: CD31 signals confer immune privilege to the vascular endothelium

    doi: 10.1073/pnas.1509627112

    Figure Lengend Snippet: Enhanced rejection of CD31-deficient skin grafts. WT female mice received either a WT or CD31-KO male-derived skin graft. As a control, female WT and CD31-KO female-derived skin was used. ( A ) Graft survival ( n = 6). **** P

    Article Snippet: As a blocking reagent , rat anti-mouse CD31 Antibody clone 390 (eBioscience) was used at a concentration of 10 μg/mL.

    Techniques: Mouse Assay, Derivative Assay

    Characterization of an anti-CD31 blocking antibody. ( A ) Subconfluent WT and CD31-KO EC monolayers were fixed in 4% paraformaldehyde (pH 7.4) for 15 min at room temperature. Following permeabilization and blocking, the slides were coincubated with an anti-CD31

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: CD31 signals confer immune privilege to the vascular endothelium

    doi: 10.1073/pnas.1509627112

    Figure Lengend Snippet: Characterization of an anti-CD31 blocking antibody. ( A ) Subconfluent WT and CD31-KO EC monolayers were fixed in 4% paraformaldehyde (pH 7.4) for 15 min at room temperature. Following permeabilization and blocking, the slides were coincubated with an anti-CD31

    Article Snippet: As a blocking reagent , rat anti-mouse CD31 Antibody clone 390 (eBioscience) was used at a concentration of 10 μg/mL.

    Techniques: Blocking Assay

    Donor endothelium is maintained in skin allografts. WT female mice received either a WT or CD31-KO male-derived skin graft. As a control, female WT and CD31-KO female-derived skin was used. Skin grafts were excised 7 d posttransplantation and were observed

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: CD31 signals confer immune privilege to the vascular endothelium

    doi: 10.1073/pnas.1509627112

    Figure Lengend Snippet: Donor endothelium is maintained in skin allografts. WT female mice received either a WT or CD31-KO male-derived skin graft. As a control, female WT and CD31-KO female-derived skin was used. Skin grafts were excised 7 d posttransplantation and were observed

    Article Snippet: As a blocking reagent , rat anti-mouse CD31 Antibody clone 390 (eBioscience) was used at a concentration of 10 μg/mL.

    Techniques: Mouse Assay, Derivative Assay

    Apoptosis-related gene profile of WT and CD31-KO ECs before and after exposure to TNF-α. ( A and B ) Volcano plots showing the expression of apoptosis-related genes by WT ( A ) and CD31-deficient ( B ) ECs following exposure to TNF-α. Plots

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: CD31 signals confer immune privilege to the vascular endothelium

    doi: 10.1073/pnas.1509627112

    Figure Lengend Snippet: Apoptosis-related gene profile of WT and CD31-KO ECs before and after exposure to TNF-α. ( A and B ) Volcano plots showing the expression of apoptosis-related genes by WT ( A ) and CD31-deficient ( B ) ECs following exposure to TNF-α. Plots

    Article Snippet: As a blocking reagent , rat anti-mouse CD31 Antibody clone 390 (eBioscience) was used at a concentration of 10 μg/mL.

    Techniques: Expressing

    Antiapoptotic pathways activated by CD31 in ECs. ( A ) Immunoprecipitation of CD31 molecules from ECs exposed to TNF-α for 20 min followed by immunoblotting with an anti-phosphotyrosine antibody and an anti-SHP2 antibody. ( B – D ) Erk ( B and

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: CD31 signals confer immune privilege to the vascular endothelium

    doi: 10.1073/pnas.1509627112

    Figure Lengend Snippet: Antiapoptotic pathways activated by CD31 in ECs. ( A ) Immunoprecipitation of CD31 molecules from ECs exposed to TNF-α for 20 min followed by immunoblotting with an anti-phosphotyrosine antibody and an anti-SHP2 antibody. ( B – D ) Erk ( B and

    Article Snippet: As a blocking reagent , rat anti-mouse CD31 Antibody clone 390 (eBioscience) was used at a concentration of 10 μg/mL.

    Techniques: Immunoprecipitation

    Spi6 gene expression is induced by proapoptotic stimuli in pancreatic β cells independently of CD31 expression and signaling. Expression of Spi6 mRNA by WT and CD31-transduced MIN6 pancreatic β cells either untreated or exposed to TNF-α

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: CD31 signals confer immune privilege to the vascular endothelium

    doi: 10.1073/pnas.1509627112

    Figure Lengend Snippet: Spi6 gene expression is induced by proapoptotic stimuli in pancreatic β cells independently of CD31 expression and signaling. Expression of Spi6 mRNA by WT and CD31-transduced MIN6 pancreatic β cells either untreated or exposed to TNF-α

    Article Snippet: As a blocking reagent , rat anti-mouse CD31 Antibody clone 390 (eBioscience) was used at a concentration of 10 μg/mL.

    Techniques: Expressing

    Spi6 gene expression is induced by proapoptotic stimuli irrespective of CD31 signals. Mock-, WT-, CD31 Y663F -, and CD31 Y686F -transduced ECs were exposed to TNF-α (50 ng/mL) for 6 h or were left untreated. Expression of Spi6 mRNA was quantified

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: CD31 signals confer immune privilege to the vascular endothelium

    doi: 10.1073/pnas.1509627112

    Figure Lengend Snippet: Spi6 gene expression is induced by proapoptotic stimuli irrespective of CD31 signals. Mock-, WT-, CD31 Y663F -, and CD31 Y686F -transduced ECs were exposed to TNF-α (50 ng/mL) for 6 h or were left untreated. Expression of Spi6 mRNA was quantified

    Article Snippet: As a blocking reagent , rat anti-mouse CD31 Antibody clone 390 (eBioscience) was used at a concentration of 10 μg/mL.

    Techniques: Expressing

    CD31 protects from extrinsic apoptosis. ( A and B ) WT and CD31-KO ECs were exposed to TNF-α (50 ng/mL) for 6 h, and apoptosis was measured by TUNEL assay. ( C ) Alternatively, WT ECs were pretreated with either a blocking anti-CD31 or an isotype

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: CD31 signals confer immune privilege to the vascular endothelium

    doi: 10.1073/pnas.1509627112

    Figure Lengend Snippet: CD31 protects from extrinsic apoptosis. ( A and B ) WT and CD31-KO ECs were exposed to TNF-α (50 ng/mL) for 6 h, and apoptosis was measured by TUNEL assay. ( C ) Alternatively, WT ECs were pretreated with either a blocking anti-CD31 or an isotype

    Article Snippet: As a blocking reagent , rat anti-mouse CD31 Antibody clone 390 (eBioscience) was used at a concentration of 10 μg/mL.

    Techniques: TUNEL Assay, Blocking Assay

    CD31-transduced pancreatic β cells are protected from extrinsic apoptosis. ( A ) Surface expression of CD31 by mock- and CD31-transfected MIN6 cells. ( B ) Insulin secretion by untreated and TNF-α–exposed mock- and CD31-transfected

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: CD31 signals confer immune privilege to the vascular endothelium

    doi: 10.1073/pnas.1509627112

    Figure Lengend Snippet: CD31-transduced pancreatic β cells are protected from extrinsic apoptosis. ( A ) Surface expression of CD31 by mock- and CD31-transfected MIN6 cells. ( B ) Insulin secretion by untreated and TNF-α–exposed mock- and CD31-transfected

    Article Snippet: As a blocking reagent , rat anti-mouse CD31 Antibody clone 390 (eBioscience) was used at a concentration of 10 μg/mL.

    Techniques: Expressing, Transfection

    The antiapoptotic gene cFlar is instrumental to CD31-mediated cytoprotection. mRNA was isolated from CD31-KO or WT ECs either untreated or stimulated with TNF-α (50 ng/mL) for 6 h. ( A and B ) cFlar mRNA quantification ( A ) and protein expression

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: CD31 signals confer immune privilege to the vascular endothelium

    doi: 10.1073/pnas.1509627112

    Figure Lengend Snippet: The antiapoptotic gene cFlar is instrumental to CD31-mediated cytoprotection. mRNA was isolated from CD31-KO or WT ECs either untreated or stimulated with TNF-α (50 ng/mL) for 6 h. ( A and B ) cFlar mRNA quantification ( A ) and protein expression

    Article Snippet: As a blocking reagent , rat anti-mouse CD31 Antibody clone 390 (eBioscience) was used at a concentration of 10 μg/mL.

    Techniques: Isolation, Expressing

    Both CD31 ITIMs are required for its prosurvival activity. CD31 gene constructs with mutations leading to the loss-of-function amino acid substitutions Y663F and Y686F in the ITIMs were generated and expressed by lentiviral transduction into CD31-KO ECs

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: CD31 signals confer immune privilege to the vascular endothelium

    doi: 10.1073/pnas.1509627112

    Figure Lengend Snippet: Both CD31 ITIMs are required for its prosurvival activity. CD31 gene constructs with mutations leading to the loss-of-function amino acid substitutions Y663F and Y686F in the ITIMs were generated and expressed by lentiviral transduction into CD31-KO ECs

    Article Snippet: As a blocking reagent , rat anti-mouse CD31 Antibody clone 390 (eBioscience) was used at a concentration of 10 μg/mL.

    Techniques: Activity Assay, Construct, Generated, Transduction

    CD31 does not affect the expression of PDL-1 or Spi6 or antioxidant activity by ECs. ( A ) Surface expression of the inhibitory coreceptor PDL-1 by untreated or TNF-α–treated WT and CD31-KO ECs was assessed by flow cytometry. ( B ) Expression

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: CD31 signals confer immune privilege to the vascular endothelium

    doi: 10.1073/pnas.1509627112

    Figure Lengend Snippet: CD31 does not affect the expression of PDL-1 or Spi6 or antioxidant activity by ECs. ( A ) Surface expression of the inhibitory coreceptor PDL-1 by untreated or TNF-α–treated WT and CD31-KO ECs was assessed by flow cytometry. ( B ) Expression

    Article Snippet: As a blocking reagent , rat anti-mouse CD31 Antibody clone 390 (eBioscience) was used at a concentration of 10 μg/mL.

    Techniques: Expressing, Antioxidant Activity Assay, Flow Cytometry, Cytometry

    CD31-induced prosurvival pathway. (1) Upon DR triggering with their cognate ligands (DR-L), the DISC forms and initiates the extrinsic pathway of apoptosis; also, the transcription factor FoxO3 is induced to translocate into the nucleus where it promotes

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: CD31 signals confer immune privilege to the vascular endothelium

    doi: 10.1073/pnas.1509627112

    Figure Lengend Snippet: CD31-induced prosurvival pathway. (1) Upon DR triggering with their cognate ligands (DR-L), the DISC forms and initiates the extrinsic pathway of apoptosis; also, the transcription factor FoxO3 is induced to translocate into the nucleus where it promotes

    Article Snippet: As a blocking reagent , rat anti-mouse CD31 Antibody clone 390 (eBioscience) was used at a concentration of 10 μg/mL.

    Techniques:

    CD31-expressing pancreatic β cells become resistant to T-cell–mediated allograft rejection. ( A and B ) The function of mock-transduced and CD31 MIN6 islets was evaluated by measuring blood glucose levels in grafted syngeneic (C57BL/6) (

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: CD31 signals confer immune privilege to the vascular endothelium

    doi: 10.1073/pnas.1509627112

    Figure Lengend Snippet: CD31-expressing pancreatic β cells become resistant to T-cell–mediated allograft rejection. ( A and B ) The function of mock-transduced and CD31 MIN6 islets was evaluated by measuring blood glucose levels in grafted syngeneic (C57BL/6) (

    Article Snippet: As a blocking reagent , rat anti-mouse CD31 Antibody clone 390 (eBioscience) was used at a concentration of 10 μg/mL.

    Techniques: Expressing

    CD31 modulates the proapoptotic transcriptional program downstream of DR signaling. ( A – C ) Expression of Fas mRNA and surface receptor by WT and CD31-KO ECs either untreated or exposed to TNF-α for 6 h as quantified by qRT-PCR ( A ) and flow

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: CD31 signals confer immune privilege to the vascular endothelium

    doi: 10.1073/pnas.1509627112

    Figure Lengend Snippet: CD31 modulates the proapoptotic transcriptional program downstream of DR signaling. ( A – C ) Expression of Fas mRNA and surface receptor by WT and CD31-KO ECs either untreated or exposed to TNF-α for 6 h as quantified by qRT-PCR ( A ) and flow

    Article Snippet: As a blocking reagent , rat anti-mouse CD31 Antibody clone 390 (eBioscience) was used at a concentration of 10 μg/mL.

    Techniques: Expressing, Quantitative RT-PCR, Flow Cytometry

    Antiapoptotic pathways activated in CD31-transduced pancreatic β cells. ( A – F ) Erk and Akt activation ( A ), Fas surface expression ( B ), caspase 7 expression and activity ( C and D ), caspase 8 activity ( E ), and cFlar protein expression ( F

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: CD31 signals confer immune privilege to the vascular endothelium

    doi: 10.1073/pnas.1509627112

    Figure Lengend Snippet: Antiapoptotic pathways activated in CD31-transduced pancreatic β cells. ( A – F ) Erk and Akt activation ( A ), Fas surface expression ( B ), caspase 7 expression and activity ( C and D ), caspase 8 activity ( E ), and cFlar protein expression ( F

    Article Snippet: As a blocking reagent , rat anti-mouse CD31 Antibody clone 390 (eBioscience) was used at a concentration of 10 μg/mL.

    Techniques: Activation Assay, Expressing, Activity Assay

    Skin phenotype of Aspn -/- mice. (A) Asporin immunoblotting of dorsal skin extracts from three wild-type and three Aspn -/- mice. (B) 2 month-old female dorsal skins were analyzed histologically by staining with Massson Trichrome, or (C) by immunohistochemistry by staining for the presence of CD31-positive blood vessels and F4/80-positive macrophages; bars in A and B are 100 μm. Similar results were obtained from analyses of six mice from each genotype. (D) Transmission electron microscopy on cross-sectioned collagen fibrils in reticular dermis. (E) Transmission electron microscopy on longitudinally oriented collagen fibrils in reticular dermis. Bars in C and D are 200 nm. (F) Quantification of collagen fibril diameter in reticular dermis. 1,000 fibrils were measured from electron microscopy images collected from six wild-type and six Aspn -/- mice.

    Journal: PLoS ONE

    Article Title: Asporin-deficient mice have tougher skin and altered skin glycosaminoglycan content and structure

    doi: 10.1371/journal.pone.0184028

    Figure Lengend Snippet: Skin phenotype of Aspn -/- mice. (A) Asporin immunoblotting of dorsal skin extracts from three wild-type and three Aspn -/- mice. (B) 2 month-old female dorsal skins were analyzed histologically by staining with Massson Trichrome, or (C) by immunohistochemistry by staining for the presence of CD31-positive blood vessels and F4/80-positive macrophages; bars in A and B are 100 μm. Similar results were obtained from analyses of six mice from each genotype. (D) Transmission electron microscopy on cross-sectioned collagen fibrils in reticular dermis. (E) Transmission electron microscopy on longitudinally oriented collagen fibrils in reticular dermis. Bars in C and D are 200 nm. (F) Quantification of collagen fibril diameter in reticular dermis. 1,000 fibrils were measured from electron microscopy images collected from six wild-type and six Aspn -/- mice.

    Article Snippet: Sections were stained with Masson Trichrome (Sigma) or processed for immunohistochemistry: rehydrated sections were washed 2 x 5 min in Tris-buffered saline (TBS) plus 0.025% Triton X-100 and blocked with 10% goat serum and 1% bovine serum albumin (BSA) for 2 h. Sections were then incubated for 16 h at 4°C with rat anti-mouse CD31 (Optistain, clone SZ31) or rat anti-mouse F4/80 (BioRad, clone CI:A3-1) diluted to 10 μg/mL in 1% BSA in TBS.

    Techniques: Mouse Assay, Staining, Immunohistochemistry, Transmission Assay, Electron Microscopy

    Overexpression of miR‐21 abrogates dihydromyricetin (DMY)‐increased DDAH1 expression and endothelial nitric oxide (NO) synthase phosphorylation and reduced ADMA levels. HUVECs were transfected with 100 n m agomir NC or miR‐21 agomir for 24 h and then treated with ox‐LDL (120 μg/mL) or ox‐LDL plus DMY (25 μmol/L) for 16 h and harvested for indicated experiments. A Western blot analysis of DDAH1 and p‐endothelial NO synthase (eNOS; ser1177) expression. n = 3 independent experiments. B, ELISA analysis of ADMA levels in cell lysates. n = 3 independent experiment. C, D, Representative images and quantification show DDAH1 (C) and phosphor‐eNOS (ser1177) (D) expression in endothelial cells in the aortic sinus lesions from DMY combined with NS‐m injection or DMY combined with 21‐m injection‐treated Apoe − / − mice fed with HFD for 12 wk. Frozen sections of aortic sinus were stained for anti‐DDAH1 or p‐eNOS (green), anti‐CD31 (red) and DAPI (blue). The dashed line area indicates differential DDAH1 or phosphor‐eNOS expression in endothelial cells. Scale: 50 μm. n = 7 mice per group. E, Western blot analysis of DDAH1 and p‐eNOS (ser1177) expression in liver. n = 7 mice per group. F and G, ELISA analysis of ADMA levels in plasma (F) and livers (G) from DMY combined with NS‐m injection or DMY combined with 21‐m injection‐treated Apoe − / − mice fed with HFD for 12 wk. n = 7 mice per group. Data shown are mean ± SEM. * P

    Journal: Journal of Cellular and Molecular Medicine

    Article Title: Dihydromyricetin increases endothelial nitric oxide production and inhibits atherosclerosis through microRNA‐21 in apolipoprotein E‐deficient mice, et al. Dihydromyricetin increases endothelial nitric oxide production and inhibits atherosclerosis through microRNA‐21 in apolipoprotein E‐deficient mice

    doi: 10.1111/jcmm.15278

    Figure Lengend Snippet: Overexpression of miR‐21 abrogates dihydromyricetin (DMY)‐increased DDAH1 expression and endothelial nitric oxide (NO) synthase phosphorylation and reduced ADMA levels. HUVECs were transfected with 100 n m agomir NC or miR‐21 agomir for 24 h and then treated with ox‐LDL (120 μg/mL) or ox‐LDL plus DMY (25 μmol/L) for 16 h and harvested for indicated experiments. A Western blot analysis of DDAH1 and p‐endothelial NO synthase (eNOS; ser1177) expression. n = 3 independent experiments. B, ELISA analysis of ADMA levels in cell lysates. n = 3 independent experiment. C, D, Representative images and quantification show DDAH1 (C) and phosphor‐eNOS (ser1177) (D) expression in endothelial cells in the aortic sinus lesions from DMY combined with NS‐m injection or DMY combined with 21‐m injection‐treated Apoe − / − mice fed with HFD for 12 wk. Frozen sections of aortic sinus were stained for anti‐DDAH1 or p‐eNOS (green), anti‐CD31 (red) and DAPI (blue). The dashed line area indicates differential DDAH1 or phosphor‐eNOS expression in endothelial cells. Scale: 50 μm. n = 7 mice per group. E, Western blot analysis of DDAH1 and p‐eNOS (ser1177) expression in liver. n = 7 mice per group. F and G, ELISA analysis of ADMA levels in plasma (F) and livers (G) from DMY combined with NS‐m injection or DMY combined with 21‐m injection‐treated Apoe − / − mice fed with HFD for 12 wk. n = 7 mice per group. Data shown are mean ± SEM. * P

    Article Snippet: For immunofluorescence staining, aortic root sections were stained with rat anti‐Mac‐2 (1:100, CL8942AP; Cedarlane), FITC‐labelled anti‐actin, α‐smooth muscle (α‐SMA; 1:500, F3777; Sigma), rat anti‐CD31 (1:200, ab23864; Abcam), rabbit anti‐VCAM1 (1:250, ab134047; Abcam), rabbit anti‐p‐eNOS (ser1177; 1:250, ab184154; Abcam) and rabbit anti‐DDAH1 (1:250, ab180599; Abcam) for 3 hours at room temperature, followed by incubation with Alexa Fluor 555 (1:300, A21434; Invitrogen) or 488 (1:300, A11034; Invitrogen) labelled secondary antibody for 1 hour at room temperature.

    Techniques: Over Expression, Expressing, Transfection, Western Blot, Enzyme-linked Immunosorbent Assay, Injection, Mouse Assay, Staining

    Dihydromyricetin (DMY) administration increases DDAH1/endothelial nitric oxide synthase (eNOS) activation in vivo and in vitro. A, B, Representative images and quantification show DDAH1 (A) and phosphor‐eNOS (ser1177) (B) expression in endothelial cells in the aortic sinus lesions rom vehicle control, DMY‐treated Apoe − / − mice fed with HFD for 12 wk. Frozen sections of aortic sinus were stained for anti‐DDAH1 or phosphor‐eNOS (green), anti‐CD31 (red) and 4′,6‐diamidino‐2‐phenylindole (DAPI; blue). The dashed line area indicates differential DDAH1 or phosphor‐eNOS expression in endothelial cells. Scale: 50 μm. n = 8‐10 mice per group. C, Western blot analysis of DDAH1 and p‐eNOS (ser1177) expression in liver. n = 8‐10 mice per group. D, E, ELISA analysis of ADMA levels in plasma (D) and livers (E) from vehicle control, DMY‐treated Apoe − / − mice fed with HFD for 12 wk. n = 8‐10 mice per group. HUVECs were treated with ox‐LDL (120 μg/mL) or ox‐LDL plus DMY (25 μmol/L) for 16 hours and harvested for indicated experiments. F, Western blot analysis of DDAH1 and p‐eNOS (ser1177) expression. n = 3 independent experiments. G, ELISA analysis of ADMA levels in cell lysates. n = 3 independent experiments. Data shown are mean ± SEM. * P

    Journal: Journal of Cellular and Molecular Medicine

    Article Title: Dihydromyricetin increases endothelial nitric oxide production and inhibits atherosclerosis through microRNA‐21 in apolipoprotein E‐deficient mice, et al. Dihydromyricetin increases endothelial nitric oxide production and inhibits atherosclerosis through microRNA‐21 in apolipoprotein E‐deficient mice

    doi: 10.1111/jcmm.15278

    Figure Lengend Snippet: Dihydromyricetin (DMY) administration increases DDAH1/endothelial nitric oxide synthase (eNOS) activation in vivo and in vitro. A, B, Representative images and quantification show DDAH1 (A) and phosphor‐eNOS (ser1177) (B) expression in endothelial cells in the aortic sinus lesions rom vehicle control, DMY‐treated Apoe − / − mice fed with HFD for 12 wk. Frozen sections of aortic sinus were stained for anti‐DDAH1 or phosphor‐eNOS (green), anti‐CD31 (red) and 4′,6‐diamidino‐2‐phenylindole (DAPI; blue). The dashed line area indicates differential DDAH1 or phosphor‐eNOS expression in endothelial cells. Scale: 50 μm. n = 8‐10 mice per group. C, Western blot analysis of DDAH1 and p‐eNOS (ser1177) expression in liver. n = 8‐10 mice per group. D, E, ELISA analysis of ADMA levels in plasma (D) and livers (E) from vehicle control, DMY‐treated Apoe − / − mice fed with HFD for 12 wk. n = 8‐10 mice per group. HUVECs were treated with ox‐LDL (120 μg/mL) or ox‐LDL plus DMY (25 μmol/L) for 16 hours and harvested for indicated experiments. F, Western blot analysis of DDAH1 and p‐eNOS (ser1177) expression. n = 3 independent experiments. G, ELISA analysis of ADMA levels in cell lysates. n = 3 independent experiments. Data shown are mean ± SEM. * P

    Article Snippet: For immunofluorescence staining, aortic root sections were stained with rat anti‐Mac‐2 (1:100, CL8942AP; Cedarlane), FITC‐labelled anti‐actin, α‐smooth muscle (α‐SMA; 1:500, F3777; Sigma), rat anti‐CD31 (1:200, ab23864; Abcam), rabbit anti‐VCAM1 (1:250, ab134047; Abcam), rabbit anti‐p‐eNOS (ser1177; 1:250, ab184154; Abcam) and rabbit anti‐DDAH1 (1:250, ab180599; Abcam) for 3 hours at room temperature, followed by incubation with Alexa Fluor 555 (1:300, A21434; Invitrogen) or 488 (1:300, A11034; Invitrogen) labelled secondary antibody for 1 hour at room temperature.

    Techniques: Activation Assay, In Vivo, In Vitro, Expressing, Mouse Assay, Staining, Western Blot, Enzyme-linked Immunosorbent Assay

    Systemic delivery of miR‐21 mimics blocks the protective effect of dihydromyricetin (DMY) on atherosclerosis in Apoe − / − mice. A, Schema of experimental procedure. B, Lesion areas were quantified by Oil Red O (ORO)‐stained thoracoabdominal aorta. C, Lesion areas were quantified by ORO‐stained aortic sinus sections. Scale: 200 μm. D, E, Representative images and quantification show Mac‐2–positive macrophages (D) and CD4‐positive T cells (E) in the aortic sinus lesions. Scale: 200 μm (D) and 100 μm (E). Arrows indicate CD4‐positve T cells in aortic sinus lesions. F, Representative images and quantification show α‐smooth muscle actin (α‐SMA)‐positive smooth muscle cells (SMCs) accumulation in the aortic sinus lesions. Arrows indicate differential SMCs accumulation in aortic sinus lesions. Scale: 200 μm. G, ELISA analysis of nitric oxide levels in plasma from DMY combined with NS‐m injection or DMY combined with 21‐m injection‐treated Apoe − / − mice. H, Representative images and quantification show VCAM‐1 expression in endothelial cells in the aortic sinus lesions. Frozen sections of aortic sinus were stained for anti‐VCAM‐1 (green), anti‐CD31 (red) and 4′,6‐diamidino‐2‐phenylindole (DAPI; blue). The dashed line area indicates differential VCAM‐1 expression in endothelial cells. Scale: 100 μm. Data shown are mean ± SEM (n = 7 mice per group). * P

    Journal: Journal of Cellular and Molecular Medicine

    Article Title: Dihydromyricetin increases endothelial nitric oxide production and inhibits atherosclerosis through microRNA‐21 in apolipoprotein E‐deficient mice, et al. Dihydromyricetin increases endothelial nitric oxide production and inhibits atherosclerosis through microRNA‐21 in apolipoprotein E‐deficient mice

    doi: 10.1111/jcmm.15278

    Figure Lengend Snippet: Systemic delivery of miR‐21 mimics blocks the protective effect of dihydromyricetin (DMY) on atherosclerosis in Apoe − / − mice. A, Schema of experimental procedure. B, Lesion areas were quantified by Oil Red O (ORO)‐stained thoracoabdominal aorta. C, Lesion areas were quantified by ORO‐stained aortic sinus sections. Scale: 200 μm. D, E, Representative images and quantification show Mac‐2–positive macrophages (D) and CD4‐positive T cells (E) in the aortic sinus lesions. Scale: 200 μm (D) and 100 μm (E). Arrows indicate CD4‐positve T cells in aortic sinus lesions. F, Representative images and quantification show α‐smooth muscle actin (α‐SMA)‐positive smooth muscle cells (SMCs) accumulation in the aortic sinus lesions. Arrows indicate differential SMCs accumulation in aortic sinus lesions. Scale: 200 μm. G, ELISA analysis of nitric oxide levels in plasma from DMY combined with NS‐m injection or DMY combined with 21‐m injection‐treated Apoe − / − mice. H, Representative images and quantification show VCAM‐1 expression in endothelial cells in the aortic sinus lesions. Frozen sections of aortic sinus were stained for anti‐VCAM‐1 (green), anti‐CD31 (red) and 4′,6‐diamidino‐2‐phenylindole (DAPI; blue). The dashed line area indicates differential VCAM‐1 expression in endothelial cells. Scale: 100 μm. Data shown are mean ± SEM (n = 7 mice per group). * P

    Article Snippet: For immunofluorescence staining, aortic root sections were stained with rat anti‐Mac‐2 (1:100, CL8942AP; Cedarlane), FITC‐labelled anti‐actin, α‐smooth muscle (α‐SMA; 1:500, F3777; Sigma), rat anti‐CD31 (1:200, ab23864; Abcam), rabbit anti‐VCAM1 (1:250, ab134047; Abcam), rabbit anti‐p‐eNOS (ser1177; 1:250, ab184154; Abcam) and rabbit anti‐DDAH1 (1:250, ab180599; Abcam) for 3 hours at room temperature, followed by incubation with Alexa Fluor 555 (1:300, A21434; Invitrogen) or 488 (1:300, A11034; Invitrogen) labelled secondary antibody for 1 hour at room temperature.

    Techniques: Mouse Assay, Staining, Enzyme-linked Immunosorbent Assay, Injection, Expressing

    Inhibition of nitric oxide (NO) production by L‐NAME abrogates the inhibitory effects of dihydromyricetin (DMY) on atherosclerosis in Apoe − / − mice. A, Lesion areas were quantified by Oil Red O (ORO)‐stained thoracoabdominal aorta. B, Lesion areas were quantified by ORO‐stained aortic sinus sections. Scale: 200 μm. C, D, Representative images and quantification show Mac‐2–positive macrophages (C) and CD4‐positive T cells (D) in the aortic sinus lesions. Scale: 200 μm (C) and 100 μm (D). Arrows indicate CD4‐positve T cells in aortic sinus lesions. E, Representative images and quantification show α‐smooth muscle actin (α‐SMA)‐positive smooth muscle cells (SMCs) accumulation in the aortic sinus lesions. Arrows indicate differential SMCs accumulation in aortic sinus lesions. Scale: 200 μm. F, ELISA analysis of NO levels in plasma from vehicle, DMY or DMY combined with L‐NAME–treated Apoe − / − mice. G, Representative images and quantification show VCAM‐1 expression in endothelial cells in the aortic sinus lesions. Frozen sections of aortic sinus were stained for anti‐VCAM‐1 (green), anti‐CD31 (red) and 4′,6‐diamidino‐2‐phenylindole (DAPI; blue). The dashed line area indicates differential VCAM‐1 expression in endothelial cells. Scale: 100 μm. Data shown are mean ± SEM (n = 7‐10 mice per group). * P

    Journal: Journal of Cellular and Molecular Medicine

    Article Title: Dihydromyricetin increases endothelial nitric oxide production and inhibits atherosclerosis through microRNA‐21 in apolipoprotein E‐deficient mice, et al. Dihydromyricetin increases endothelial nitric oxide production and inhibits atherosclerosis through microRNA‐21 in apolipoprotein E‐deficient mice

    doi: 10.1111/jcmm.15278

    Figure Lengend Snippet: Inhibition of nitric oxide (NO) production by L‐NAME abrogates the inhibitory effects of dihydromyricetin (DMY) on atherosclerosis in Apoe − / − mice. A, Lesion areas were quantified by Oil Red O (ORO)‐stained thoracoabdominal aorta. B, Lesion areas were quantified by ORO‐stained aortic sinus sections. Scale: 200 μm. C, D, Representative images and quantification show Mac‐2–positive macrophages (C) and CD4‐positive T cells (D) in the aortic sinus lesions. Scale: 200 μm (C) and 100 μm (D). Arrows indicate CD4‐positve T cells in aortic sinus lesions. E, Representative images and quantification show α‐smooth muscle actin (α‐SMA)‐positive smooth muscle cells (SMCs) accumulation in the aortic sinus lesions. Arrows indicate differential SMCs accumulation in aortic sinus lesions. Scale: 200 μm. F, ELISA analysis of NO levels in plasma from vehicle, DMY or DMY combined with L‐NAME–treated Apoe − / − mice. G, Representative images and quantification show VCAM‐1 expression in endothelial cells in the aortic sinus lesions. Frozen sections of aortic sinus were stained for anti‐VCAM‐1 (green), anti‐CD31 (red) and 4′,6‐diamidino‐2‐phenylindole (DAPI; blue). The dashed line area indicates differential VCAM‐1 expression in endothelial cells. Scale: 100 μm. Data shown are mean ± SEM (n = 7‐10 mice per group). * P

    Article Snippet: For immunofluorescence staining, aortic root sections were stained with rat anti‐Mac‐2 (1:100, CL8942AP; Cedarlane), FITC‐labelled anti‐actin, α‐smooth muscle (α‐SMA; 1:500, F3777; Sigma), rat anti‐CD31 (1:200, ab23864; Abcam), rabbit anti‐VCAM1 (1:250, ab134047; Abcam), rabbit anti‐p‐eNOS (ser1177; 1:250, ab184154; Abcam) and rabbit anti‐DDAH1 (1:250, ab180599; Abcam) for 3 hours at room temperature, followed by incubation with Alexa Fluor 555 (1:300, A21434; Invitrogen) or 488 (1:300, A11034; Invitrogen) labelled secondary antibody for 1 hour at room temperature.

    Techniques: Inhibition, Mouse Assay, Staining, Enzyme-linked Immunosorbent Assay, Expressing

    Effect of AMI on rat SVF cells. a Cell size (μm) of the SVF cells, demonstrating no difference between the three groups. b Percentage of colony-forming cells, showing significantly fewer colonies in the 1D group (6.1 ± 1.6 %) compared to the Control (11.4 ± 1.8 %) and 7D groups (11.0 ± 0.9 %). c Number of cells, obtained 4 days after seeding of the SVF fraction, demonstrating significantly fewer cells in the 1D group (1.3 ± 0.1 million cells) compared to the Control (2.0 ± 0.1 million cells) and 7D groups (1.8 ± 0.1 million cells). d Percentage of positive cells for the markers CD31, CD34, CD45, CD73, CD90, CD105 and CD271 showing significantly fewer CD90- and CD105-positive cells in the 1D group. Data shown as mean ± SD (Control and 7D group n = 6, 1D group n = 5, * p

    Journal: Cell and Tissue Research

    Article Title: Acute myocardial infarction does not affect functional characteristics of adipose-derived stem cells in rats, but reduces the number of stem cells in adipose tissue

    doi: 10.1007/s00441-015-2239-z

    Figure Lengend Snippet: Effect of AMI on rat SVF cells. a Cell size (μm) of the SVF cells, demonstrating no difference between the three groups. b Percentage of colony-forming cells, showing significantly fewer colonies in the 1D group (6.1 ± 1.6 %) compared to the Control (11.4 ± 1.8 %) and 7D groups (11.0 ± 0.9 %). c Number of cells, obtained 4 days after seeding of the SVF fraction, demonstrating significantly fewer cells in the 1D group (1.3 ± 0.1 million cells) compared to the Control (2.0 ± 0.1 million cells) and 7D groups (1.8 ± 0.1 million cells). d Percentage of positive cells for the markers CD31, CD34, CD45, CD73, CD90, CD105 and CD271 showing significantly fewer CD90- and CD105-positive cells in the 1D group. Data shown as mean ± SD (Control and 7D group n = 6, 1D group n = 5, * p

    Article Snippet: The following primary antibodies were used: phycoerythrin (PE)-conjugated mouse-α-rat antibodies against CD90 (1:20; Cedarlane Laboratories, Ontario, Canada), CD45 (1:20; BD Biosciences, San José, CA, USA), CD271 (1:25; BD Biosciences) and CD31 (1:20; AbD Serotec, Oxford, UK), PE-labeled mouse α-human antibody CD105 (1:25; Caltag, Invitrogen, USA), as well as unlabeled mouse-α-rat antibodies against CD34 (1:20; Santa Cruz Biotechnology) and CD73 (1:20; BD Biosciences).

    Techniques:

    Effect of AMI on cultured rat ASC. a Representative light microscopy image demonstrating the morphology of cultured rat ASC. b Cell size (μm) of ASC, demonstrating no significant differences between the three groups. c Population doubling time (in days) depicted for passages 1–3, showing no significant differences between the three groups. d Percentage of positive cells for the markers CD31, CD34, CD45, CD73, CD90, CD105 and CD271, showing no significant differences in numbers of positive cells between the groups. Data shown as mean ± SD (Control and 7D group n = 6, 1D group n = 5)

    Journal: Cell and Tissue Research

    Article Title: Acute myocardial infarction does not affect functional characteristics of adipose-derived stem cells in rats, but reduces the number of stem cells in adipose tissue

    doi: 10.1007/s00441-015-2239-z

    Figure Lengend Snippet: Effect of AMI on cultured rat ASC. a Representative light microscopy image demonstrating the morphology of cultured rat ASC. b Cell size (μm) of ASC, demonstrating no significant differences between the three groups. c Population doubling time (in days) depicted for passages 1–3, showing no significant differences between the three groups. d Percentage of positive cells for the markers CD31, CD34, CD45, CD73, CD90, CD105 and CD271, showing no significant differences in numbers of positive cells between the groups. Data shown as mean ± SD (Control and 7D group n = 6, 1D group n = 5)

    Article Snippet: The following primary antibodies were used: phycoerythrin (PE)-conjugated mouse-α-rat antibodies against CD90 (1:20; Cedarlane Laboratories, Ontario, Canada), CD45 (1:20; BD Biosciences, San José, CA, USA), CD271 (1:25; BD Biosciences) and CD31 (1:20; AbD Serotec, Oxford, UK), PE-labeled mouse α-human antibody CD105 (1:25; Caltag, Invitrogen, USA), as well as unlabeled mouse-α-rat antibodies against CD34 (1:20; Santa Cruz Biotechnology) and CD73 (1:20; BD Biosciences).

    Techniques: Cell Culture, Light Microscopy