cd54 Search Results


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R&D Systems recombinant human icam
Cell density of endothelial cells cultivated in the presence of soluble <t>CD54.</t> Endothelial cells cultivated in the presence of soluble CD54 (sCD54) were expanded for five splits. After the first (p1), second (p2) and fifth (p5) passages (as shown in the x-axis) the cellular densities were measured (y-axis) and compared between different growing conditions: in the presence of sCD54 alone, resulting in higher propagation rate, than cell culture treated with HUVEC-conditioned medium collected at p4 (CMP4) alone.
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Sino Biological icam1
Mechanical Rigidity of <t>ICAM1</t> Nano-Topographies Controls T Cell On-Ridge Spreading and In-Groove Invasiveness Plasticity Balance (A) 3D super-resolution reconstruction of the ICAM1-functionalized PAA nano-topographic surface (G’=16 kPa). (B) Test super-resolution imaging (3D reconstruction) of human CD4+ T cells spreading and migrating along the ICAM1-coated nano-topographic surfaces on soft (16 kPa) and rigid (50 kPa) PAA surfaces. (C) Schematic of the T cell morphometric analysis and metrics: T cell height, projected area of spreading of entire cell interface ( S entire cell IF ) and projected area of in-groove invasive T cell interface ( S invasive IF ). (D and E) Mechanoregulation of T cell height, spreading area and invasiveness as indicated by T cell spreading assay on soft (G’=16 kPa) and rigid (G’=50 kPa) ICAM1 nano-textures. T cell spreading enhances on the rigid ICAM1, accompanied with T cell flattening, i.e. decrease of the T cell height. Results indicate a mechanically controlled dynamic balance between on-ridge T cell spreading and in-groove invasiveness, as shown on the schematic panel (E). I.e. T cell in-groove invasiveness structurally competes with on-ridge spreading, indicating that on-ridge spreading is mechanically enhanced and out-balances in-groove invasiveness on the rigid (G’=50 kPa) ICAM1 nano-topography. Alternatively, soft (G’=16 kPa) ICAM-1 nano-textures are unable to promote the mechanically sensitive on-ridge T cell spreading, shifting the balance towards steric in-groove T cell invasiveness. Data on the plots on (D) are as follows: boxes - means, Q1 and Q3; whiskers - max and min, X - medians; p values - one way ANOVA test. Experimental data collected in triplicates, total n>50.
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Bio-Rad cd54
Fig. 2. Cell surface MHC and adhesion molecules are downregulated in SV5 V-protein-expressing LCLs. Cell surface expression of MHC class I and II molecules, <t>CD54</t> and CD19 was measured on IARC-171 and IB4 (pBabe and V-protein LCL). A representative flow cytometric analysis is shown in (a). Dark grey shading represents cell surface staining on pBabe LCLs; no shading with black outline represents cell surface staining on V-protein LCLs; light grey shading represents background FITC fluorescence obtained on cells stained with control antibodies. (b, c) A summary of MHC class I and II, CD54 and CD19 expression on IARC-1 (b) and IB4 (c) cell lines. Each result represents the percentage of cell surface expression relative to the pBabe LCL from at least three separate experiments. Error bars, SD. A paired t-test was used ascertain the statistical significance of the results; *P,0.05. (d) Total lysates from IARC-171 and IB4 (pBabe and V-protein LCL) were analysed by Western blotting using antibodies specific to MHC class I, STAT1, IRF-1, TAP1, TAP2, low-molecular weight protein-2 and actin.
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Bio-Rad cd54 antibody
Fig. 2. Cell surface MHC and adhesion molecules are downregulated in SV5 V-protein-expressing LCLs. Cell surface expression of MHC class I and II molecules, <t>CD54</t> and CD19 was measured on IARC-171 and IB4 (pBabe and V-protein LCL). A representative flow cytometric analysis is shown in (a). Dark grey shading represents cell surface staining on pBabe LCLs; no shading with black outline represents cell surface staining on V-protein LCLs; light grey shading represents background FITC fluorescence obtained on cells stained with control antibodies. (b, c) A summary of MHC class I and II, CD54 and CD19 expression on IARC-1 (b) and IB4 (c) cell lines. Each result represents the percentage of cell surface expression relative to the pBabe LCL from at least three separate experiments. Error bars, SD. A paired t-test was used ascertain the statistical significance of the results; *P,0.05. (d) Total lysates from IARC-171 and IB4 (pBabe and V-protein LCL) were analysed by Western blotting using antibodies specific to MHC class I, STAT1, IRF-1, TAP1, TAP2, low-molecular weight protein-2 and actin.
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Bio-Rad mouse anti rat icam 1
Fig. 2. Cell surface MHC and adhesion molecules are downregulated in SV5 V-protein-expressing LCLs. Cell surface expression of MHC class I and II molecules, <t>CD54</t> and CD19 was measured on IARC-171 and IB4 (pBabe and V-protein LCL). A representative flow cytometric analysis is shown in (a). Dark grey shading represents cell surface staining on pBabe LCLs; no shading with black outline represents cell surface staining on V-protein LCLs; light grey shading represents background FITC fluorescence obtained on cells stained with control antibodies. (b, c) A summary of MHC class I and II, CD54 and CD19 expression on IARC-1 (b) and IB4 (c) cell lines. Each result represents the percentage of cell surface expression relative to the pBabe LCL from at least three separate experiments. Error bars, SD. A paired t-test was used ascertain the statistical significance of the results; *P,0.05. (d) Total lysates from IARC-171 and IB4 (pBabe and V-protein LCL) were analysed by Western blotting using antibodies specific to MHC class I, STAT1, IRF-1, TAP1, TAP2, low-molecular weight protein-2 and actin.
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R&D Systems recombinant mouse icam
Fig. 2. Cell surface MHC and adhesion molecules are downregulated in SV5 V-protein-expressing LCLs. Cell surface expression of MHC class I and II molecules, <t>CD54</t> and CD19 was measured on IARC-171 and IB4 (pBabe and V-protein LCL). A representative flow cytometric analysis is shown in (a). Dark grey shading represents cell surface staining on pBabe LCLs; no shading with black outline represents cell surface staining on V-protein LCLs; light grey shading represents background FITC fluorescence obtained on cells stained with control antibodies. (b, c) A summary of MHC class I and II, CD54 and CD19 expression on IARC-1 (b) and IB4 (c) cell lines. Each result represents the percentage of cell surface expression relative to the pBabe LCL from at least three separate experiments. Error bars, SD. A paired t-test was used ascertain the statistical significance of the results; *P,0.05. (d) Total lysates from IARC-171 and IB4 (pBabe and V-protein LCL) were analysed by Western blotting using antibodies specific to MHC class I, STAT1, IRF-1, TAP1, TAP2, low-molecular weight protein-2 and actin.
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R&D Systems elisa
Fig. 2. Cell surface MHC and adhesion molecules are downregulated in SV5 V-protein-expressing LCLs. Cell surface expression of MHC class I and II molecules, <t>CD54</t> and CD19 was measured on IARC-171 and IB4 (pBabe and V-protein LCL). A representative flow cytometric analysis is shown in (a). Dark grey shading represents cell surface staining on pBabe LCLs; no shading with black outline represents cell surface staining on V-protein LCLs; light grey shading represents background FITC fluorescence obtained on cells stained with control antibodies. (b, c) A summary of MHC class I and II, CD54 and CD19 expression on IARC-1 (b) and IB4 (c) cell lines. Each result represents the percentage of cell surface expression relative to the pBabe LCL from at least three separate experiments. Error bars, SD. A paired t-test was used ascertain the statistical significance of the results; *P,0.05. (d) Total lysates from IARC-171 and IB4 (pBabe and V-protein LCL) were analysed by Western blotting using antibodies specific to MHC class I, STAT1, IRF-1, TAP1, TAP2, low-molecular weight protein-2 and actin.
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R&D Systems standard elisa kit
Fig. 2. Cell surface MHC and adhesion molecules are downregulated in SV5 V-protein-expressing LCLs. Cell surface expression of MHC class I and II molecules, <t>CD54</t> and CD19 was measured on IARC-171 and IB4 (pBabe and V-protein LCL). A representative flow cytometric analysis is shown in (a). Dark grey shading represents cell surface staining on pBabe LCLs; no shading with black outline represents cell surface staining on V-protein LCLs; light grey shading represents background FITC fluorescence obtained on cells stained with control antibodies. (b, c) A summary of MHC class I and II, CD54 and CD19 expression on IARC-1 (b) and IB4 (c) cell lines. Each result represents the percentage of cell surface expression relative to the pBabe LCL from at least three separate experiments. Error bars, SD. A paired t-test was used ascertain the statistical significance of the results; *P,0.05. (d) Total lysates from IARC-171 and IB4 (pBabe and V-protein LCL) were analysed by Western blotting using antibodies specific to MHC class I, STAT1, IRF-1, TAP1, TAP2, low-molecular weight protein-2 and actin.
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Boster Bio mouse monoclonal anti ty tag antibody
Fig. 2. Cell surface MHC and adhesion molecules are downregulated in SV5 V-protein-expressing LCLs. Cell surface expression of MHC class I and II molecules, <t>CD54</t> and CD19 was measured on IARC-171 and IB4 (pBabe and V-protein LCL). A representative flow cytometric analysis is shown in (a). Dark grey shading represents cell surface staining on pBabe LCLs; no shading with black outline represents cell surface staining on V-protein LCLs; light grey shading represents background FITC fluorescence obtained on cells stained with control antibodies. (b, c) A summary of MHC class I and II, CD54 and CD19 expression on IARC-1 (b) and IB4 (c) cell lines. Each result represents the percentage of cell surface expression relative to the pBabe LCL from at least three separate experiments. Error bars, SD. A paired t-test was used ascertain the statistical significance of the results; *P,0.05. (d) Total lysates from IARC-171 and IB4 (pBabe and V-protein LCL) were analysed by Western blotting using antibodies specific to MHC class I, STAT1, IRF-1, TAP1, TAP2, low-molecular weight protein-2 and actin.
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Cell Signaling Technology Inc anti western blot
Fig. 2. Cell surface MHC and adhesion molecules are downregulated in SV5 V-protein-expressing LCLs. Cell surface expression of MHC class I and II molecules, <t>CD54</t> and CD19 was measured on IARC-171 and IB4 (pBabe and V-protein LCL). A representative flow cytometric analysis is shown in (a). Dark grey shading represents cell surface staining on pBabe LCLs; no shading with black outline represents cell surface staining on V-protein LCLs; light grey shading represents background FITC fluorescence obtained on cells stained with control antibodies. (b, c) A summary of MHC class I and II, CD54 and CD19 expression on IARC-1 (b) and IB4 (c) cell lines. Each result represents the percentage of cell surface expression relative to the pBabe LCL from at least three separate experiments. Error bars, SD. A paired t-test was used ascertain the statistical significance of the results; *P,0.05. (d) Total lysates from IARC-171 and IB4 (pBabe and V-protein LCL) were analysed by Western blotting using antibodies specific to MHC class I, STAT1, IRF-1, TAP1, TAP2, low-molecular weight protein-2 and actin.
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Cell Signaling Technology Inc icam 1
Fig. 2. Cell surface MHC and adhesion molecules are downregulated in SV5 V-protein-expressing LCLs. Cell surface expression of MHC class I and II molecules, <t>CD54</t> and CD19 was measured on IARC-171 and IB4 (pBabe and V-protein LCL). A representative flow cytometric analysis is shown in (a). Dark grey shading represents cell surface staining on pBabe LCLs; no shading with black outline represents cell surface staining on V-protein LCLs; light grey shading represents background FITC fluorescence obtained on cells stained with control antibodies. (b, c) A summary of MHC class I and II, CD54 and CD19 expression on IARC-1 (b) and IB4 (c) cell lines. Each result represents the percentage of cell surface expression relative to the pBabe LCL from at least three separate experiments. Error bars, SD. A paired t-test was used ascertain the statistical significance of the results; *P,0.05. (d) Total lysates from IARC-171 and IB4 (pBabe and V-protein LCL) were analysed by Western blotting using antibodies specific to MHC class I, STAT1, IRF-1, TAP1, TAP2, low-molecular weight protein-2 and actin.
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R&D Systems nephrin
(A) Immunostaining with a mouse monoclonal anti human CD80 (red) was performed on paraffin embedded sections from patients with MCD and FSGS. Positive and negative controls are shown (upper panel). Upregulation of glomerular CD80 was noted in most of MCD but few FSGS patients (lower panel). (B) CD80 (green, upper panel) colocalized <t>with</t> <t>caveolin</t> 1 in an endothelial specific pattern (arrows). To less extent, CD80 (red, lower panel) colocalized with <t>nephrin</t> (arrowheads). (C) Specificity of immunostaining was confirmed by silver staining. No signal detected in kidney tissue sample from same MCD patient in relapse when primary antibody was omitted. (D) Electron microscopy (EM) from kidney tissue from patient with MCD is shown. CD80 colocalized with glomerular endothelial cells and rarely podocytes. Original EM images captured at 6000x (far right) and 30000x. Podocyte (P), capillary lumen (CL), endothelial cell (EC), urinary space (U), glomerular basement membrane (GBM). Scale bars: 100 μm.
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Image Search Results


Cell density of endothelial cells cultivated in the presence of soluble CD54. Endothelial cells cultivated in the presence of soluble CD54 (sCD54) were expanded for five splits. After the first (p1), second (p2) and fifth (p5) passages (as shown in the x-axis) the cellular densities were measured (y-axis) and compared between different growing conditions: in the presence of sCD54 alone, resulting in higher propagation rate, than cell culture treated with HUVEC-conditioned medium collected at p4 (CMP4) alone.

Journal: International Journal of Cardiology. Heart & Vasculature

Article Title: Soluble CD54 induces human endothelial cells ex vivo expansion useful for cardiovascular regeneration and tissue engineering application

doi: 10.1016/j.ijcha.2015.01.004

Figure Lengend Snippet: Cell density of endothelial cells cultivated in the presence of soluble CD54. Endothelial cells cultivated in the presence of soluble CD54 (sCD54) were expanded for five splits. After the first (p1), second (p2) and fifth (p5) passages (as shown in the x-axis) the cellular densities were measured (y-axis) and compared between different growing conditions: in the presence of sCD54 alone, resulting in higher propagation rate, than cell culture treated with HUVEC-conditioned medium collected at p4 (CMP4) alone.

Article Snippet: Recombinant human ICAM-1/CD54 (murine myeloma cell line, NSO-derived; Accession #CAA30051), CD146, KDR and CD31 were purchased from R&D Systems, Inc.; CD45 was purchased from AbD Serotec, CD34 and CD14 were purchased from Becton Dickinson Pharmingen, and fibronectin was purchased from Southern Biotech.

Techniques: Cell Culture

Capillary density. Tube formation occurred through an ordered sequence of events and was investigated with an inverted optical light microscope; (A) cells beginning to migrate and align themselves to close polygons beginning (B) to form complete tubules (C) of endothelial cells cultivated with the addition of sCD54. The histogram shows the data mean ± SD of the quantitative analysis of tube formation area; in the x-axis was reported capillary density in the presence of HUVEC-conditioned medium collected at p4 (CMP4), sCD54 and antibody direct against CD54 at the concentration of 15 ng/ml (y-axis). Scale bars: 200 μm.

Journal: International Journal of Cardiology. Heart & Vasculature

Article Title: Soluble CD54 induces human endothelial cells ex vivo expansion useful for cardiovascular regeneration and tissue engineering application

doi: 10.1016/j.ijcha.2015.01.004

Figure Lengend Snippet: Capillary density. Tube formation occurred through an ordered sequence of events and was investigated with an inverted optical light microscope; (A) cells beginning to migrate and align themselves to close polygons beginning (B) to form complete tubules (C) of endothelial cells cultivated with the addition of sCD54. The histogram shows the data mean ± SD of the quantitative analysis of tube formation area; in the x-axis was reported capillary density in the presence of HUVEC-conditioned medium collected at p4 (CMP4), sCD54 and antibody direct against CD54 at the concentration of 15 ng/ml (y-axis). Scale bars: 200 μm.

Article Snippet: Recombinant human ICAM-1/CD54 (murine myeloma cell line, NSO-derived; Accession #CAA30051), CD146, KDR and CD31 were purchased from R&D Systems, Inc.; CD45 was purchased from AbD Serotec, CD34 and CD14 were purchased from Becton Dickinson Pharmingen, and fibronectin was purchased from Southern Biotech.

Techniques: Sequencing, Light Microscopy, Concentration Assay

Mechanical Rigidity of ICAM1 Nano-Topographies Controls T Cell On-Ridge Spreading and In-Groove Invasiveness Plasticity Balance (A) 3D super-resolution reconstruction of the ICAM1-functionalized PAA nano-topographic surface (G’=16 kPa). (B) Test super-resolution imaging (3D reconstruction) of human CD4+ T cells spreading and migrating along the ICAM1-coated nano-topographic surfaces on soft (16 kPa) and rigid (50 kPa) PAA surfaces. (C) Schematic of the T cell morphometric analysis and metrics: T cell height, projected area of spreading of entire cell interface ( S entire cell IF ) and projected area of in-groove invasive T cell interface ( S invasive IF ). (D and E) Mechanoregulation of T cell height, spreading area and invasiveness as indicated by T cell spreading assay on soft (G’=16 kPa) and rigid (G’=50 kPa) ICAM1 nano-textures. T cell spreading enhances on the rigid ICAM1, accompanied with T cell flattening, i.e. decrease of the T cell height. Results indicate a mechanically controlled dynamic balance between on-ridge T cell spreading and in-groove invasiveness, as shown on the schematic panel (E). I.e. T cell in-groove invasiveness structurally competes with on-ridge spreading, indicating that on-ridge spreading is mechanically enhanced and out-balances in-groove invasiveness on the rigid (G’=50 kPa) ICAM1 nano-topography. Alternatively, soft (G’=16 kPa) ICAM-1 nano-textures are unable to promote the mechanically sensitive on-ridge T cell spreading, shifting the balance towards steric in-groove T cell invasiveness. Data on the plots on (D) are as follows: boxes - means, Q1 and Q3; whiskers - max and min, X - medians; p values - one way ANOVA test. Experimental data collected in triplicates, total n>50.

Journal: STAR Protocols

Article Title: Engineering Elastic Nano- and Micro-Patterns and Textures for Directed Cell Motility

doi: 10.1016/j.xpro.2019.100013

Figure Lengend Snippet: Mechanical Rigidity of ICAM1 Nano-Topographies Controls T Cell On-Ridge Spreading and In-Groove Invasiveness Plasticity Balance (A) 3D super-resolution reconstruction of the ICAM1-functionalized PAA nano-topographic surface (G’=16 kPa). (B) Test super-resolution imaging (3D reconstruction) of human CD4+ T cells spreading and migrating along the ICAM1-coated nano-topographic surfaces on soft (16 kPa) and rigid (50 kPa) PAA surfaces. (C) Schematic of the T cell morphometric analysis and metrics: T cell height, projected area of spreading of entire cell interface ( S entire cell IF ) and projected area of in-groove invasive T cell interface ( S invasive IF ). (D and E) Mechanoregulation of T cell height, spreading area and invasiveness as indicated by T cell spreading assay on soft (G’=16 kPa) and rigid (G’=50 kPa) ICAM1 nano-textures. T cell spreading enhances on the rigid ICAM1, accompanied with T cell flattening, i.e. decrease of the T cell height. Results indicate a mechanically controlled dynamic balance between on-ridge T cell spreading and in-groove invasiveness, as shown on the schematic panel (E). I.e. T cell in-groove invasiveness structurally competes with on-ridge spreading, indicating that on-ridge spreading is mechanically enhanced and out-balances in-groove invasiveness on the rigid (G’=50 kPa) ICAM1 nano-topography. Alternatively, soft (G’=16 kPa) ICAM-1 nano-textures are unable to promote the mechanically sensitive on-ridge T cell spreading, shifting the balance towards steric in-groove T cell invasiveness. Data on the plots on (D) are as follows: boxes - means, Q1 and Q3; whiskers - max and min, X - medians; p values - one way ANOVA test. Experimental data collected in triplicates, total n>50.

Article Snippet: ICAM1, Human Protein, Recombinant, hIgG1-Fc.His Tag, Active , Sino Biological, China , Cat#10346-H03H-50.

Techniques: Imaging

Journal: STAR Protocols

Article Title: Engineering Elastic Nano- and Micro-Patterns and Textures for Directed Cell Motility

doi: 10.1016/j.xpro.2019.100013

Figure Lengend Snippet:

Article Snippet: ICAM1, Human Protein, Recombinant, hIgG1-Fc.His Tag, Active , Sino Biological, China , Cat#10346-H03H-50.

Techniques: Recombinant, Electrophoresis, Concentration Assay, Labeling, Saline, Modification, Cell Isolation, Derivative Assay, Software

Fig. 2. Cell surface MHC and adhesion molecules are downregulated in SV5 V-protein-expressing LCLs. Cell surface expression of MHC class I and II molecules, CD54 and CD19 was measured on IARC-171 and IB4 (pBabe and V-protein LCL). A representative flow cytometric analysis is shown in (a). Dark grey shading represents cell surface staining on pBabe LCLs; no shading with black outline represents cell surface staining on V-protein LCLs; light grey shading represents background FITC fluorescence obtained on cells stained with control antibodies. (b, c) A summary of MHC class I and II, CD54 and CD19 expression on IARC-1 (b) and IB4 (c) cell lines. Each result represents the percentage of cell surface expression relative to the pBabe LCL from at least three separate experiments. Error bars, SD. A paired t-test was used ascertain the statistical significance of the results; *P,0.05. (d) Total lysates from IARC-171 and IB4 (pBabe and V-protein LCL) were analysed by Western blotting using antibodies specific to MHC class I, STAT1, IRF-1, TAP1, TAP2, low-molecular weight protein-2 and actin.

Journal: The Journal of general virology

Article Title: STAT1 contributes to the maintenance of the latency III viral programme observed in Epstein-Barr virus-transformed B cells and their recognition by CD8+ T cells.

doi: 10.1099/vir.0.011627-0

Figure Lengend Snippet: Fig. 2. Cell surface MHC and adhesion molecules are downregulated in SV5 V-protein-expressing LCLs. Cell surface expression of MHC class I and II molecules, CD54 and CD19 was measured on IARC-171 and IB4 (pBabe and V-protein LCL). A representative flow cytometric analysis is shown in (a). Dark grey shading represents cell surface staining on pBabe LCLs; no shading with black outline represents cell surface staining on V-protein LCLs; light grey shading represents background FITC fluorescence obtained on cells stained with control antibodies. (b, c) A summary of MHC class I and II, CD54 and CD19 expression on IARC-1 (b) and IB4 (c) cell lines. Each result represents the percentage of cell surface expression relative to the pBabe LCL from at least three separate experiments. Error bars, SD. A paired t-test was used ascertain the statistical significance of the results; *P,0.05. (d) Total lysates from IARC-171 and IB4 (pBabe and V-protein LCL) were analysed by Western blotting using antibodies specific to MHC class I, STAT1, IRF-1, TAP1, TAP2, low-molecular weight protein-2 and actin.

Article Snippet: Fluorescein isothiocyanate (FITC)-conjugated mAbs specific for CD54 (MCA1615F) and CD19 (MCA1940F) were purchased from Serotec.

Techniques: Expressing, Staining, Fluorescence, Control, Western Blot, Molecular Weight

(A) Immunostaining with a mouse monoclonal anti human CD80 (red) was performed on paraffin embedded sections from patients with MCD and FSGS. Positive and negative controls are shown (upper panel). Upregulation of glomerular CD80 was noted in most of MCD but few FSGS patients (lower panel). (B) CD80 (green, upper panel) colocalized with caveolin 1 in an endothelial specific pattern (arrows). To less extent, CD80 (red, lower panel) colocalized with nephrin (arrowheads). (C) Specificity of immunostaining was confirmed by silver staining. No signal detected in kidney tissue sample from same MCD patient in relapse when primary antibody was omitted. (D) Electron microscopy (EM) from kidney tissue from patient with MCD is shown. CD80 colocalized with glomerular endothelial cells and rarely podocytes. Original EM images captured at 6000x (far right) and 30000x. Podocyte (P), capillary lumen (CL), endothelial cell (EC), urinary space (U), glomerular basement membrane (GBM). Scale bars: 100 μm.

Journal: Pediatric nephrology (Berlin, Germany)

Article Title: Glomerular endothelial cells and podocytes can express CD80 in patients with minimal change disease during relapse

doi: 10.1007/s00467-020-04541-3

Figure Lengend Snippet: (A) Immunostaining with a mouse monoclonal anti human CD80 (red) was performed on paraffin embedded sections from patients with MCD and FSGS. Positive and negative controls are shown (upper panel). Upregulation of glomerular CD80 was noted in most of MCD but few FSGS patients (lower panel). (B) CD80 (green, upper panel) colocalized with caveolin 1 in an endothelial specific pattern (arrows). To less extent, CD80 (red, lower panel) colocalized with nephrin (arrowheads). (C) Specificity of immunostaining was confirmed by silver staining. No signal detected in kidney tissue sample from same MCD patient in relapse when primary antibody was omitted. (D) Electron microscopy (EM) from kidney tissue from patient with MCD is shown. CD80 colocalized with glomerular endothelial cells and rarely podocytes. Original EM images captured at 6000x (far right) and 30000x. Podocyte (P), capillary lumen (CL), endothelial cell (EC), urinary space (U), glomerular basement membrane (GBM). Scale bars: 100 μm.

Article Snippet: Reagents and antibodies The following primary antibodies were used CD80 (AF740, R&D) at 1:100 dilution; CD80 (MAB140, clone 37711, R&D) at 1:20; caveolin 1 (D46G3, #3267, Cell Signaling) at 1:800, nephrin (provided by Verma R), ICAM-1 (AF796, R&D) at 1:1000, synaptopodin (10R-2373, Fitzgerald) at 1:20.

Techniques: Immunostaining, Silver Staining, Electron Microscopy