rabbit anti-collagen iv Search Results


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  • 94
    Thermo Fisher rabbit anti collagen iv
    Rabbit Anti Collagen Iv, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore rabbit anti collagen iv
    Schematic diagram showing the mechanisms involved in the renoprotective effects of PPARδ in diabetic nephropathy. The <t>anti-inflammatory</t> transcriptional repressor Bcl-6 represses the expression of MCP-1. PPARδ activation by GW0742 releases Bcl-6, which is associated with suppression of MCP-1, to attenuate macrophage infiltration, inflammatory gene expression, and <t>type</t> <t>IV</t> <t>collagen</t> accumulation in the kidney.
    Rabbit Anti Collagen Iv, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 219 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti collagen iv/product/Millipore
    Average 99 stars, based on 219 article reviews
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    99
    Abcam anti collagen iv antibody
    Two-dimensional image analysis. A. Areas sampled for 2D quantification are shown in white across structures in a coronal section of the mouse brain. Scale bar, 500 μm. B. ROI in the cortical gray matter showing microvasculature labeled with <t>anti-collagen</t> <t>IV</t> (green), neurons labeled with <t>anti-NeuN</t> (red) and Dapi to reveal all cell nuclei (blue). Cell nuclei appearing inside green-labeled blood vessels are considered capillary-associated nuclei; cell nuclei double-labeled in red are considered neuronal cell nuclei; all remaining nuclei are identified by exclusion as glial cell nuclei. C. A Cavalieri estimator grid of 4 μm spacing (shown) was applied to images acquired under 20x magnification, and a 2μm grid was applied to images acquired under 63x magnification. The points marked in red correspond to the area of the image occupied by capillaries. Scale bar, 10μm D . Superposition of FITC-dextran (green, lumen) and collagen IV (red, basal lamina) labeling of capillaries. Single image acquired under 63x magnification. Scale bar, 10μm.
    Anti Collagen Iv Antibody, supplied by Abcam, used in various techniques. Bioz Stars score: 99/100, based on 342 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti collagen iv antibody/product/Abcam
    Average 99 stars, based on 342 article reviews
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    99
    Abcam rabbit anti collagen iv
    L.m specific memory CD8 + T cells preferentially traffic in the bloodstream. (A) 200 naïve GFP + OT-I cells were transferred in C57BL/6 mice. One day later, recipient mice were injected i.v. with 0.1×LD 50 Wt L.m -OVA. Thirty days after this immunization, recipient mice were injected i.v with 3×10 6 naïve CMTPX-labelled <t>polyclonal</t> CD8 + T cells. The following day, mice were injected i.p with PBS or 100 µg of FTY720. Animals were killed 24 hours later and their LNs, spleen and blood harvested, stained for CD8 expression and analyzed by flow cytometry. Numbers indicate the percentages of GFP + OT-I memory cells and CMTPX naive polyclonal CD8 + T cells among total CD8 + T cells. (B) Wt BALB/c mice were injected i.v with 0.1×LD 50 Wt L.m and further treated as in (A) with the exception that polyclonal naive CD8 + T cells were labelled with CFSE and that L.m specific endogenous memory CD8 + T cells were identified as LLO 91 – 99 /H-2K d tetramers + CD8 + cells. Data are representative of 3 independent experiments.
    Rabbit Anti Collagen Iv, supplied by Abcam, used in various techniques. Bioz Stars score: 99/100, based on 454 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti collagen iv/product/Abcam
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    98
    Bio-Rad rabbit anti mouse collagen iv
    T lymphocytes express ligand αvβ3 integrin and adhere to EGFL7. a Colocalization of CD4+ cells (green) and EGFL7 (red) in perivascular ECM area in MS brain lesions (upper panel = frontal, lower panel = temporal). <t>Collagen</t> <t>IV</t> = white. Nuclei (DAPI) = blue. Representative of n = 4 MS donors, ≥ 3 lesions/donor. b Expression of αvβ3 integrin on human CD14 monocytes ex vivo (left), memory CD4 T lymphocytes ex vivo (center), and in vitro activated memory CD4 T lymphocytes (right), as assessed by flow cytometry. Representative of n = 6 donors. c Expression of αvβ3 integrin on human CD4 T lymphocytes ex vivo and upon activation. Percentage and mean fluorescence intensity (MFI). n = 6 donors. d Expression of αvβ3 integrin on human CD4 T lymphocytes ex vivo from healthy controls, untreated MS or treated MS. Flow cytometry analysis. 1 dot = 1 donor. e , f Expression of αvβ3 integrin by murine CD4 T lymphocytes ( e ) ex vivo and on day 1–5 in vitro upon activation, and ( f ) in naïve and MOG 35–55 -immunized (EAE) mice at different time points of EAE. n = 4–5 mice/condition, 1 dot = 1 <t>mouse.</t> g – k Adhesion assay. g Representative microscopy images of CFSE-labeled activated human CD4 T lymphocytes adhering to rhEGFL7 or BSA 1% after 24 h. Scale bar = 100 μm. Amount of human ( h ) and murine ( j ) activated CFSE-labeled CD4 T cells adhering to coated rh or rmEGFL7, ICAM-1 or fibronectin, normalized to BSA 1%, according to fluorescence intensity as assessed by plate-reader. n = 6 human donors and n = 8 mice. Amount of human ( i ) and murine ( k ) cells adhering to coated EGFL7, ICAM-1 or fibronectin, following pre-treatment with rhEGFL7 ( i ) or rmEGFL7 ( k ); <t>anti-αvβ3</t> integrin neutralizing antibody ( i ) or RGD-peptide ( k ); or relevant control (rat serum, isotype or RAD-peptide). n = 7 human donors and n = 8 mice. Statistical analysis performed by Wilcoxon matched-pairs signed rank test ( c ) and one-way ANOVA followed by Tukey’s multiple comparison test ( d – k ). * p
    Rabbit Anti Mouse Collagen Iv, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 98/100, based on 167 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 98 stars, based on 167 article reviews
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    89
    Abcam rabbit anti mouse collagen iv
    PECAM and CD99 function past the level of the endothelium in C57BL/6 mice. A : wild-type FVB/n and C57BL/6 mice were pretreated with control IgG, <t>anti-PECAM,</t> or anti-CD99 (3 mg/kg) prior to ear stimulation with croton oil (1% solution, 20 μl/ear) or carrier alone. After 4 h, ear tissue was collected and immunohistochemical staining was performed for PECAM [endothelial cells (ECs), red], myeloid-related protein 14 (neutrophils, green), and <t>collagen</t> <t>IV</t> [basement membrane (BM), purple]. 3-Dimensional confocal images were acquired from each sample and subsequently analyzed. Insets : z -orthogonal view along the indicated line to demonstrate leukocyte arrest in relation to the endothelium and basement membrane at 1.5 times the xy image. Scale bar = 50 μm. B : quantification of findings in A (percent leukocyte extravasation within 50 μm of postcapillary venules in FVB/n mice). C : schematic of quantification of site of leukocyte blockade. Leukocytes were scored as being in 1 of 6 positions: 1 ) luminal, 2 ) apically arrested, 3 ) arrested partway through the endothelium, 4 ) arrested at the level of the basement membrane, 5 ) actively migrating through the basement membrane, or 6 ) fully extravasated. D : quantification of the site of leukocyte arrest in FVB/n mice. E : quantification of findings in A (percent leukocyte extravasation within 50 μm of postcapillary venules in C57BL/6 mice). F : quantification of the site of leukocyte arrest in C57BL/6 mice. G : schematic model of anti-PECAM and anti-CD99 arrest in FVB/n and C57BL/6 mice. One hundred to 200 cells were analyzed per ear. Carrier-alone stimulation resulted in minimal leukocyte recruitment (data not shown). Total leukocytes per field of view, vessel length, and vessel diameter were equivalent for all treatment groups (data not shown). Data are representative of recordings from ≥4 mice per group with ≥8 images quantified per <t>mouse.</t> * P
    Rabbit Anti Mouse Collagen Iv, supplied by Abcam, used in various techniques. Bioz Stars score: 89/100, based on 35 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Cosmo Bio rabbit anti mouse type iv collagen polyclonal antibody
    Treadmill, median survival times, and histological study. (a) All CBA recipients of cardiac allografts exercised on a treadmill system, (b) cardiac graft survival in CBA recipients of a B6 heart that were treated on a treadmill for 1 week after grafting, 1 week before grafting, and 1 week before and after grafting and untreated allograft recipients. (c) Histologic examination (hematoxylin–eosin (HE) staining) of cardiac allografts obtained 2 and 4 weeks after grafting from postoperative 1-week treadmill-exercised recipients and untreated recipients, and assessment of HE staining. (d, e) Results of double immunostaining of cardiac allografts obtained 4 weeks after transplantation from postoperative 1-week treadmill-exercised mice and untreated mice. Fresh 4-μm-thick graft cryosections were incubated with anti-CD4, CD8, and CD68 monoclonal and anti-Foxp3 <t>polyclonal</t> antibody (magnification ×10 or ×100). MST median survival time. Ex Tx exercise treatment. No Tx no treatment. * P
    Rabbit Anti Mouse Type Iv Collagen Polyclonal Antibody, supplied by Cosmo Bio, used in various techniques. Bioz Stars score: 88/100, based on 179 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    Millipore anti collagen iv antibody
    Treadmill, median survival times, and histological study. (a) All CBA recipients of cardiac allografts exercised on a treadmill system, (b) cardiac graft survival in CBA recipients of a B6 heart that were treated on a treadmill for 1 week after grafting, 1 week before grafting, and 1 week before and after grafting and untreated allograft recipients. (c) Histologic examination (hematoxylin–eosin (HE) staining) of cardiac allografts obtained 2 and 4 weeks after grafting from postoperative 1-week treadmill-exercised recipients and untreated recipients, and assessment of HE staining. (d, e) Results of double immunostaining of cardiac allografts obtained 4 weeks after transplantation from postoperative 1-week treadmill-exercised mice and untreated mice. Fresh 4-μm-thick graft cryosections were incubated with anti-CD4, CD8, and CD68 monoclonal and anti-Foxp3 <t>polyclonal</t> antibody (magnification ×10 or ×100). MST median survival time. Ex Tx exercise treatment. No Tx no treatment. * P
    Anti Collagen Iv Antibody, supplied by Millipore, used in various techniques. Bioz Stars score: 95/100, based on 52 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Abcam rabbit polyclonal anti collagen type iv
    Treadmill, median survival times, and histological study. (a) All CBA recipients of cardiac allografts exercised on a treadmill system, (b) cardiac graft survival in CBA recipients of a B6 heart that were treated on a treadmill for 1 week after grafting, 1 week before grafting, and 1 week before and after grafting and untreated allograft recipients. (c) Histologic examination (hematoxylin–eosin (HE) staining) of cardiac allografts obtained 2 and 4 weeks after grafting from postoperative 1-week treadmill-exercised recipients and untreated recipients, and assessment of HE staining. (d, e) Results of double immunostaining of cardiac allografts obtained 4 weeks after transplantation from postoperative 1-week treadmill-exercised mice and untreated mice. Fresh 4-μm-thick graft cryosections were incubated with anti-CD4, CD8, and CD68 monoclonal and anti-Foxp3 <t>polyclonal</t> antibody (magnification ×10 or ×100). MST median survival time. Ex Tx exercise treatment. No Tx no treatment. * P
    Rabbit Polyclonal Anti Collagen Type Iv, supplied by Abcam, used in various techniques. Bioz Stars score: 89/100, based on 44 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Rockland Immunochemicals anti collagen type iv rabbit antibody 600 401 106 0 5
    Treadmill, median survival times, and histological study. (a) All CBA recipients of cardiac allografts exercised on a treadmill system, (b) cardiac graft survival in CBA recipients of a B6 heart that were treated on a treadmill for 1 week after grafting, 1 week before grafting, and 1 week before and after grafting and untreated allograft recipients. (c) Histologic examination (hematoxylin–eosin (HE) staining) of cardiac allografts obtained 2 and 4 weeks after grafting from postoperative 1-week treadmill-exercised recipients and untreated recipients, and assessment of HE staining. (d, e) Results of double immunostaining of cardiac allografts obtained 4 weeks after transplantation from postoperative 1-week treadmill-exercised mice and untreated mice. Fresh 4-μm-thick graft cryosections were incubated with anti-CD4, CD8, and CD68 monoclonal and anti-Foxp3 <t>polyclonal</t> antibody (magnification ×10 or ×100). MST median survival time. Ex Tx exercise treatment. No Tx no treatment. * P
    Anti Collagen Type Iv Rabbit Antibody 600 401 106 0 5, supplied by Rockland Immunochemicals, used in various techniques. Bioz Stars score: 94/100, based on 19 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti collagen type iv rabbit antibody 600 401 106 0 5/product/Rockland Immunochemicals
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    96
    Abcam rabbit anti type iv collagen polyclonal antibody
    Treadmill, median survival times, and histological study. (a) All CBA recipients of cardiac allografts exercised on a treadmill system, (b) cardiac graft survival in CBA recipients of a B6 heart that were treated on a treadmill for 1 week after grafting, 1 week before grafting, and 1 week before and after grafting and untreated allograft recipients. (c) Histologic examination (hematoxylin–eosin (HE) staining) of cardiac allografts obtained 2 and 4 weeks after grafting from postoperative 1-week treadmill-exercised recipients and untreated recipients, and assessment of HE staining. (d, e) Results of double immunostaining of cardiac allografts obtained 4 weeks after transplantation from postoperative 1-week treadmill-exercised mice and untreated mice. Fresh 4-μm-thick graft cryosections were incubated with anti-CD4, CD8, and CD68 monoclonal and anti-Foxp3 <t>polyclonal</t> antibody (magnification ×10 or ×100). MST median survival time. Ex Tx exercise treatment. No Tx no treatment. * P
    Rabbit Anti Type Iv Collagen Polyclonal Antibody, supplied by Abcam, used in various techniques. Bioz Stars score: 96/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Schematic diagram showing the mechanisms involved in the renoprotective effects of PPARδ in diabetic nephropathy. The anti-inflammatory transcriptional repressor Bcl-6 represses the expression of MCP-1. PPARδ activation by GW0742 releases Bcl-6, which is associated with suppression of MCP-1, to attenuate macrophage infiltration, inflammatory gene expression, and type IV collagen accumulation in the kidney.

    Journal: Diabetes

    Article Title: Activation of Peroxisome Proliferator-Activated Receptor ? Inhibits Streptozotocin-Induced Diabetic Nephropathy Through Anti-Inflammatory Mechanisms in Mice

    doi: 10.2337/db10-1361

    Figure Lengend Snippet: Schematic diagram showing the mechanisms involved in the renoprotective effects of PPARδ in diabetic nephropathy. The anti-inflammatory transcriptional repressor Bcl-6 represses the expression of MCP-1. PPARδ activation by GW0742 releases Bcl-6, which is associated with suppression of MCP-1, to attenuate macrophage infiltration, inflammatory gene expression, and type IV collagen accumulation in the kidney.

    Article Snippet: To clarify the differences in mesangial matrix proteins, we used rabbit anti-type IV collagen antibody (Millipore, Temecula, CA), followed by Alexa Fluor 488 goat anti-rabbit IgG (Invitrogen, Carlsbad, CA).

    Techniques: Expressing, Activation Assay

    Two-dimensional image analysis. A. Areas sampled for 2D quantification are shown in white across structures in a coronal section of the mouse brain. Scale bar, 500 μm. B. ROI in the cortical gray matter showing microvasculature labeled with anti-collagen IV (green), neurons labeled with anti-NeuN (red) and Dapi to reveal all cell nuclei (blue). Cell nuclei appearing inside green-labeled blood vessels are considered capillary-associated nuclei; cell nuclei double-labeled in red are considered neuronal cell nuclei; all remaining nuclei are identified by exclusion as glial cell nuclei. C. A Cavalieri estimator grid of 4 μm spacing (shown) was applied to images acquired under 20x magnification, and a 2μm grid was applied to images acquired under 63x magnification. The points marked in red correspond to the area of the image occupied by capillaries. Scale bar, 10μm D . Superposition of FITC-dextran (green, lumen) and collagen IV (red, basal lamina) labeling of capillaries. Single image acquired under 63x magnification. Scale bar, 10μm.

    Journal: bioRxiv

    Article Title: Energy supply per neuron is constrained by capillary density in the mouse brain

    doi: 10.1101/2020.02.03.932434

    Figure Lengend Snippet: Two-dimensional image analysis. A. Areas sampled for 2D quantification are shown in white across structures in a coronal section of the mouse brain. Scale bar, 500 μm. B. ROI in the cortical gray matter showing microvasculature labeled with anti-collagen IV (green), neurons labeled with anti-NeuN (red) and Dapi to reveal all cell nuclei (blue). Cell nuclei appearing inside green-labeled blood vessels are considered capillary-associated nuclei; cell nuclei double-labeled in red are considered neuronal cell nuclei; all remaining nuclei are identified by exclusion as glial cell nuclei. C. A Cavalieri estimator grid of 4 μm spacing (shown) was applied to images acquired under 20x magnification, and a 2μm grid was applied to images acquired under 63x magnification. The points marked in red correspond to the area of the image occupied by capillaries. Scale bar, 10μm D . Superposition of FITC-dextran (green, lumen) and collagen IV (red, basal lamina) labeling of capillaries. Single image acquired under 63x magnification. Scale bar, 10μm.

    Article Snippet: Sections from animals not injected with FITC-Dextran were next incubated under stirring for 48 hours at 4°C in PB containing rabbit polyclonal anti-collagen IV antibody (Abcam, Ab6586) at a 1:500 dilution.

    Techniques: Labeling

    Muscle ECM deposition, fiber diameter, and vascular markers of the muscle-specific ILK-deficient mice. A : Immunohistochemical detection of ColIV and laminin in paraffin-embedded gastrocnemius sections ( n = 5–9). Representative images are presented at original magnification ×20. B : Values of integrated intensity of staining for ColIV and laminin are presented as mean ± SEM, and data are normalized to chow ILK lox/lox . # P

    Journal: Diabetes

    Article Title: Integrin-Linked Kinase in Muscle Is Necessary for the Development of Insulin Resistance in Diet-Induced Obese Mice

    doi: 10.2337/db15-1434

    Figure Lengend Snippet: Muscle ECM deposition, fiber diameter, and vascular markers of the muscle-specific ILK-deficient mice. A : Immunohistochemical detection of ColIV and laminin in paraffin-embedded gastrocnemius sections ( n = 5–9). Representative images are presented at original magnification ×20. B : Values of integrated intensity of staining for ColIV and laminin are presented as mean ± SEM, and data are normalized to chow ILK lox/lox . # P

    Article Snippet: Immunohistochemistry ILK, collagen IV (ColIV), laminin, CD31, and von Willebrand factor (vWF) were assessed by immunohistochemistry in paraffin-embedded gastrocnemius tissue sections (5 μm) using the following primary antibodies: anti-ILK (sc-20019, 1:150; Santa Cruz Biotechnology), anti-ColIV (ab6586, 1:100; Abcam), anti-laminin (Z0097, 1:3,000; DakoCytomation), anti-CD31 (550274, 1:200; BD Biosciences), and anti-vWF (A0082, 1:3,000; DakoCytomation).

    Techniques: Mouse Assay, Immunohistochemistry, Staining

    L.m specific memory CD8 + T cells preferentially traffic in the bloodstream. (A) 200 naïve GFP + OT-I cells were transferred in C57BL/6 mice. One day later, recipient mice were injected i.v. with 0.1×LD 50 Wt L.m -OVA. Thirty days after this immunization, recipient mice were injected i.v with 3×10 6 naïve CMTPX-labelled polyclonal CD8 + T cells. The following day, mice were injected i.p with PBS or 100 µg of FTY720. Animals were killed 24 hours later and their LNs, spleen and blood harvested, stained for CD8 expression and analyzed by flow cytometry. Numbers indicate the percentages of GFP + OT-I memory cells and CMTPX naive polyclonal CD8 + T cells among total CD8 + T cells. (B) Wt BALB/c mice were injected i.v with 0.1×LD 50 Wt L.m and further treated as in (A) with the exception that polyclonal naive CD8 + T cells were labelled with CFSE and that L.m specific endogenous memory CD8 + T cells were identified as LLO 91 – 99 /H-2K d tetramers + CD8 + cells. Data are representative of 3 independent experiments.

    Journal: PLoS ONE

    Article Title: Visualizing Early Splenic Memory CD8+ T Cells Reactivation against Intracellular Bacteria in the Mouse

    doi: 10.1371/journal.pone.0011524

    Figure Lengend Snippet: L.m specific memory CD8 + T cells preferentially traffic in the bloodstream. (A) 200 naïve GFP + OT-I cells were transferred in C57BL/6 mice. One day later, recipient mice were injected i.v. with 0.1×LD 50 Wt L.m -OVA. Thirty days after this immunization, recipient mice were injected i.v with 3×10 6 naïve CMTPX-labelled polyclonal CD8 + T cells. The following day, mice were injected i.p with PBS or 100 µg of FTY720. Animals were killed 24 hours later and their LNs, spleen and blood harvested, stained for CD8 expression and analyzed by flow cytometry. Numbers indicate the percentages of GFP + OT-I memory cells and CMTPX naive polyclonal CD8 + T cells among total CD8 + T cells. (B) Wt BALB/c mice were injected i.v with 0.1×LD 50 Wt L.m and further treated as in (A) with the exception that polyclonal naive CD8 + T cells were labelled with CFSE and that L.m specific endogenous memory CD8 + T cells were identified as LLO 91 – 99 /H-2K d tetramers + CD8 + cells. Data are representative of 3 independent experiments.

    Article Snippet: Rabbit polyclonal anti-collagen IV antibody was purchased from Abcam (Cambridge, MA).

    Techniques: Mouse Assay, Injection, Staining, Expressing, Flow Cytometry, Cytometry

    T lymphocytes express ligand αvβ3 integrin and adhere to EGFL7. a Colocalization of CD4+ cells (green) and EGFL7 (red) in perivascular ECM area in MS brain lesions (upper panel = frontal, lower panel = temporal). Collagen IV = white. Nuclei (DAPI) = blue. Representative of n = 4 MS donors, ≥ 3 lesions/donor. b Expression of αvβ3 integrin on human CD14 monocytes ex vivo (left), memory CD4 T lymphocytes ex vivo (center), and in vitro activated memory CD4 T lymphocytes (right), as assessed by flow cytometry. Representative of n = 6 donors. c Expression of αvβ3 integrin on human CD4 T lymphocytes ex vivo and upon activation. Percentage and mean fluorescence intensity (MFI). n = 6 donors. d Expression of αvβ3 integrin on human CD4 T lymphocytes ex vivo from healthy controls, untreated MS or treated MS. Flow cytometry analysis. 1 dot = 1 donor. e , f Expression of αvβ3 integrin by murine CD4 T lymphocytes ( e ) ex vivo and on day 1–5 in vitro upon activation, and ( f ) in naïve and MOG 35–55 -immunized (EAE) mice at different time points of EAE. n = 4–5 mice/condition, 1 dot = 1 mouse. g – k Adhesion assay. g Representative microscopy images of CFSE-labeled activated human CD4 T lymphocytes adhering to rhEGFL7 or BSA 1% after 24 h. Scale bar = 100 μm. Amount of human ( h ) and murine ( j ) activated CFSE-labeled CD4 T cells adhering to coated rh or rmEGFL7, ICAM-1 or fibronectin, normalized to BSA 1%, according to fluorescence intensity as assessed by plate-reader. n = 6 human donors and n = 8 mice. Amount of human ( i ) and murine ( k ) cells adhering to coated EGFL7, ICAM-1 or fibronectin, following pre-treatment with rhEGFL7 ( i ) or rmEGFL7 ( k ); anti-αvβ3 integrin neutralizing antibody ( i ) or RGD-peptide ( k ); or relevant control (rat serum, isotype or RAD-peptide). n = 7 human donors and n = 8 mice. Statistical analysis performed by Wilcoxon matched-pairs signed rank test ( c ) and one-way ANOVA followed by Tukey’s multiple comparison test ( d – k ). * p

    Journal: Nature Communications

    Article Title: EGFL7 reduces CNS inflammation in mouse

    doi: 10.1038/s41467-018-03186-z

    Figure Lengend Snippet: T lymphocytes express ligand αvβ3 integrin and adhere to EGFL7. a Colocalization of CD4+ cells (green) and EGFL7 (red) in perivascular ECM area in MS brain lesions (upper panel = frontal, lower panel = temporal). Collagen IV = white. Nuclei (DAPI) = blue. Representative of n = 4 MS donors, ≥ 3 lesions/donor. b Expression of αvβ3 integrin on human CD14 monocytes ex vivo (left), memory CD4 T lymphocytes ex vivo (center), and in vitro activated memory CD4 T lymphocytes (right), as assessed by flow cytometry. Representative of n = 6 donors. c Expression of αvβ3 integrin on human CD4 T lymphocytes ex vivo and upon activation. Percentage and mean fluorescence intensity (MFI). n = 6 donors. d Expression of αvβ3 integrin on human CD4 T lymphocytes ex vivo from healthy controls, untreated MS or treated MS. Flow cytometry analysis. 1 dot = 1 donor. e , f Expression of αvβ3 integrin by murine CD4 T lymphocytes ( e ) ex vivo and on day 1–5 in vitro upon activation, and ( f ) in naïve and MOG 35–55 -immunized (EAE) mice at different time points of EAE. n = 4–5 mice/condition, 1 dot = 1 mouse. g – k Adhesion assay. g Representative microscopy images of CFSE-labeled activated human CD4 T lymphocytes adhering to rhEGFL7 or BSA 1% after 24 h. Scale bar = 100 μm. Amount of human ( h ) and murine ( j ) activated CFSE-labeled CD4 T cells adhering to coated rh or rmEGFL7, ICAM-1 or fibronectin, normalized to BSA 1%, according to fluorescence intensity as assessed by plate-reader. n = 6 human donors and n = 8 mice. Amount of human ( i ) and murine ( k ) cells adhering to coated EGFL7, ICAM-1 or fibronectin, following pre-treatment with rhEGFL7 ( i ) or rmEGFL7 ( k ); anti-αvβ3 integrin neutralizing antibody ( i ) or RGD-peptide ( k ); or relevant control (rat serum, isotype or RAD-peptide). n = 7 human donors and n = 8 mice. Statistical analysis performed by Wilcoxon matched-pairs signed rank test ( c ) and one-way ANOVA followed by Tukey’s multiple comparison test ( d – k ). * p

    Article Snippet: The following primary antibodies were used: mouse anti-human collagen IV (Abcam; 1:200) or goat anti-human collagen IV (Millipore; 1:80), mouse anti-human CD31 (BD Biosciences; 1:200), rabbit anti-human EGFL7 (clone 57; 1:100), mouse anti-human CD4 (BD Biosciences; 1:80), rat anti-mouse CD31 (BD Biosciences; 1:500), rat anti-mouse laminin-alpha4 (RnD systems; 1:500), donkey anti-mouse IgG-AF488 (Invitrogen; 1:500), rabbit anti-mouse collagen IV (BioRad; 1:100), rat anti-mouse CD4 (BD Biosciences AF647-labeled; 1:200), rabbit anti-mouse fibrinogen (Innovative Research; 1:1500), rabbit anti-mouse CD146 (Abcam; 1:200), rat anti-mouse CD106 (eBioscience; 1:100) and rat anti-mouse CD54 (eBioscience; 1:100).

    Techniques: Mass Spectrometry, Expressing, Ex Vivo, In Vitro, Flow Cytometry, Cytometry, Activation Assay, Fluorescence, Mouse Assay, Cell Adhesion Assay, Microscopy, Labeling

    PECAM and CD99 function past the level of the endothelium in C57BL/6 mice. A : wild-type FVB/n and C57BL/6 mice were pretreated with control IgG, anti-PECAM, or anti-CD99 (3 mg/kg) prior to ear stimulation with croton oil (1% solution, 20 μl/ear) or carrier alone. After 4 h, ear tissue was collected and immunohistochemical staining was performed for PECAM [endothelial cells (ECs), red], myeloid-related protein 14 (neutrophils, green), and collagen IV [basement membrane (BM), purple]. 3-Dimensional confocal images were acquired from each sample and subsequently analyzed. Insets : z -orthogonal view along the indicated line to demonstrate leukocyte arrest in relation to the endothelium and basement membrane at 1.5 times the xy image. Scale bar = 50 μm. B : quantification of findings in A (percent leukocyte extravasation within 50 μm of postcapillary venules in FVB/n mice). C : schematic of quantification of site of leukocyte blockade. Leukocytes were scored as being in 1 of 6 positions: 1 ) luminal, 2 ) apically arrested, 3 ) arrested partway through the endothelium, 4 ) arrested at the level of the basement membrane, 5 ) actively migrating through the basement membrane, or 6 ) fully extravasated. D : quantification of the site of leukocyte arrest in FVB/n mice. E : quantification of findings in A (percent leukocyte extravasation within 50 μm of postcapillary venules in C57BL/6 mice). F : quantification of the site of leukocyte arrest in C57BL/6 mice. G : schematic model of anti-PECAM and anti-CD99 arrest in FVB/n and C57BL/6 mice. One hundred to 200 cells were analyzed per ear. Carrier-alone stimulation resulted in minimal leukocyte recruitment (data not shown). Total leukocytes per field of view, vessel length, and vessel diameter were equivalent for all treatment groups (data not shown). Data are representative of recordings from ≥4 mice per group with ≥8 images quantified per mouse. * P

    Journal: American Journal of Physiology - Heart and Circulatory Physiology

    Article Title: 4D intravital microscopy uncovers critical strain differences for the roles of PECAM and CD99 in leukocyte diapedesis

    doi: 10.1152/ajpheart.00289.2016

    Figure Lengend Snippet: PECAM and CD99 function past the level of the endothelium in C57BL/6 mice. A : wild-type FVB/n and C57BL/6 mice were pretreated with control IgG, anti-PECAM, or anti-CD99 (3 mg/kg) prior to ear stimulation with croton oil (1% solution, 20 μl/ear) or carrier alone. After 4 h, ear tissue was collected and immunohistochemical staining was performed for PECAM [endothelial cells (ECs), red], myeloid-related protein 14 (neutrophils, green), and collagen IV [basement membrane (BM), purple]. 3-Dimensional confocal images were acquired from each sample and subsequently analyzed. Insets : z -orthogonal view along the indicated line to demonstrate leukocyte arrest in relation to the endothelium and basement membrane at 1.5 times the xy image. Scale bar = 50 μm. B : quantification of findings in A (percent leukocyte extravasation within 50 μm of postcapillary venules in FVB/n mice). C : schematic of quantification of site of leukocyte blockade. Leukocytes were scored as being in 1 of 6 positions: 1 ) luminal, 2 ) apically arrested, 3 ) arrested partway through the endothelium, 4 ) arrested at the level of the basement membrane, 5 ) actively migrating through the basement membrane, or 6 ) fully extravasated. D : quantification of the site of leukocyte arrest in FVB/n mice. E : quantification of findings in A (percent leukocyte extravasation within 50 μm of postcapillary venules in C57BL/6 mice). F : quantification of the site of leukocyte arrest in C57BL/6 mice. G : schematic model of anti-PECAM and anti-CD99 arrest in FVB/n and C57BL/6 mice. One hundred to 200 cells were analyzed per ear. Carrier-alone stimulation resulted in minimal leukocyte recruitment (data not shown). Total leukocytes per field of view, vessel length, and vessel diameter were equivalent for all treatment groups (data not shown). Data are representative of recordings from ≥4 mice per group with ≥8 images quantified per mouse. * P

    Article Snippet: Rabbit anti-mouse collagen IV, rat anti-mouse myeloid-related protein 14 (MRP14), and rabbit anti-mouse α-smooth muscle actin (α-SMA) were purchased from Abcam (Cambridge, MA).

    Techniques: Mouse Assay, Immunohistochemistry, Staining

    Treadmill, median survival times, and histological study. (a) All CBA recipients of cardiac allografts exercised on a treadmill system, (b) cardiac graft survival in CBA recipients of a B6 heart that were treated on a treadmill for 1 week after grafting, 1 week before grafting, and 1 week before and after grafting and untreated allograft recipients. (c) Histologic examination (hematoxylin–eosin (HE) staining) of cardiac allografts obtained 2 and 4 weeks after grafting from postoperative 1-week treadmill-exercised recipients and untreated recipients, and assessment of HE staining. (d, e) Results of double immunostaining of cardiac allografts obtained 4 weeks after transplantation from postoperative 1-week treadmill-exercised mice and untreated mice. Fresh 4-μm-thick graft cryosections were incubated with anti-CD4, CD8, and CD68 monoclonal and anti-Foxp3 polyclonal antibody (magnification ×10 or ×100). MST median survival time. Ex Tx exercise treatment. No Tx no treatment. * P

    Journal: Transplant International

    Article Title: Treadmill exercise induces murine cardiac allograft survival and generates regulatory T cell

    doi: 10.1111/tri.12491

    Figure Lengend Snippet: Treadmill, median survival times, and histological study. (a) All CBA recipients of cardiac allografts exercised on a treadmill system, (b) cardiac graft survival in CBA recipients of a B6 heart that were treated on a treadmill for 1 week after grafting, 1 week before grafting, and 1 week before and after grafting and untreated allograft recipients. (c) Histologic examination (hematoxylin–eosin (HE) staining) of cardiac allografts obtained 2 and 4 weeks after grafting from postoperative 1-week treadmill-exercised recipients and untreated recipients, and assessment of HE staining. (d, e) Results of double immunostaining of cardiac allografts obtained 4 weeks after transplantation from postoperative 1-week treadmill-exercised mice and untreated mice. Fresh 4-μm-thick graft cryosections were incubated with anti-CD4, CD8, and CD68 monoclonal and anti-Foxp3 polyclonal antibody (magnification ×10 or ×100). MST median survival time. Ex Tx exercise treatment. No Tx no treatment. * P

    Article Snippet: Cryosections were then incubated with rabbit anti-mouse type IV collagen polyclonal antibody (LB1403; Cosmo Bio, Tokyo) and peroxidase-conjugated anti-rabbit Ig (55693; Mitsubishi Chemical, Tokyo) and then developed brown with diaminobenzidine (Vector Laboratories).

    Techniques: Crocin Bleaching Assay, Staining, Double Immunostaining, Transplantation Assay, Mouse Assay, Incubation, Microscale Thermophoresis

    Evidence of generation of regulatory cells in treadmill-exercised CBA allograft recipients. (a) Scheme on adoptive transfer study to confirm the generation of regulatory T cells. (b, c) Cardiac allograft survival after adoptive transfer of whole splenocytes (b) or CD4 + cells (c). (d–h) Results of double immunostaining of cardiac allografts obtained 4 weeks after transplantation from untreated mice and postoperative 1-week treadmill-exercised mice (d–g) and 100 days after adoptive transfer of CD4 + cell from longtime surviving secondary CBA recipients with B6 beating heart (h). Fresh 4-μm-thick graft cryosections were incubated with anti-CD4, CD8, and CD68 monoclonal antibody or anti-Foxp3 polyclonal antibody. In (d–g), the left-hand panels show samples obtained from mice exercising on a treadmill, and the right-hand panels show samples from untreated mice (magnification ×40). In (h), all panels show samples obtained from longtime surviving transplant recipients in CD4 + cell adoptive transfer groups (magnification ×100) (i) CD4, CD25, and Foxp3 expression in splenocytes as determined by flow cytometry 1, 2, and 4 weeks after transplantation. The right-hand graph shows the percentage of CD4 + CD25 + Foxp3 + cells in the CD4 + cells as determined by flow cytometry. Data are mean ± SD values ( n = 5 mice in each group). MST median survival time. * P

    Journal: Transplant International

    Article Title: Treadmill exercise induces murine cardiac allograft survival and generates regulatory T cell

    doi: 10.1111/tri.12491

    Figure Lengend Snippet: Evidence of generation of regulatory cells in treadmill-exercised CBA allograft recipients. (a) Scheme on adoptive transfer study to confirm the generation of regulatory T cells. (b, c) Cardiac allograft survival after adoptive transfer of whole splenocytes (b) or CD4 + cells (c). (d–h) Results of double immunostaining of cardiac allografts obtained 4 weeks after transplantation from untreated mice and postoperative 1-week treadmill-exercised mice (d–g) and 100 days after adoptive transfer of CD4 + cell from longtime surviving secondary CBA recipients with B6 beating heart (h). Fresh 4-μm-thick graft cryosections were incubated with anti-CD4, CD8, and CD68 monoclonal antibody or anti-Foxp3 polyclonal antibody. In (d–g), the left-hand panels show samples obtained from mice exercising on a treadmill, and the right-hand panels show samples from untreated mice (magnification ×40). In (h), all panels show samples obtained from longtime surviving transplant recipients in CD4 + cell adoptive transfer groups (magnification ×100) (i) CD4, CD25, and Foxp3 expression in splenocytes as determined by flow cytometry 1, 2, and 4 weeks after transplantation. The right-hand graph shows the percentage of CD4 + CD25 + Foxp3 + cells in the CD4 + cells as determined by flow cytometry. Data are mean ± SD values ( n = 5 mice in each group). MST median survival time. * P

    Article Snippet: Cryosections were then incubated with rabbit anti-mouse type IV collagen polyclonal antibody (LB1403; Cosmo Bio, Tokyo) and peroxidase-conjugated anti-rabbit Ig (55693; Mitsubishi Chemical, Tokyo) and then developed brown with diaminobenzidine (Vector Laboratories).

    Techniques: Crocin Bleaching Assay, Adoptive Transfer Assay, Double Immunostaining, Transplantation Assay, Mouse Assay, Incubation, Expressing, Flow Cytometry, Cytometry, Microscale Thermophoresis