Structured Review

Abcam anti cd81 antibody
Characterization of the exosomes isolated from the peritoneal dialysis effluent (PDE). (a) Morphology of the exosomes observed by a transmission electron microscopy. Scale bar = 100 nm. (b) The particle size distribution of exosomes measured by nanoparticle tracking analysis. (c) Exosomes surface markers (HSP70, CD63, and <t>CD81)</t> detected using western blot.
Anti Cd81 Antibody, supplied by Abcam, used in various techniques. Bioz Stars score: 94/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Average 94 stars, based on 11 article reviews
Price from $9.99 to $1999.99
anti cd81 antibody - by Bioz Stars, 2022-10
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Images

1) Product Images from "Differentially Expressed microRNAs in Peritoneal Dialysis Effluent-Derived Exosomes from the Patients with Ultrafiltration Failure"

Article Title: Differentially Expressed microRNAs in Peritoneal Dialysis Effluent-Derived Exosomes from the Patients with Ultrafiltration Failure

Journal: Genetics Research

doi: 10.1155/2022/2276175

Characterization of the exosomes isolated from the peritoneal dialysis effluent (PDE). (a) Morphology of the exosomes observed by a transmission electron microscopy. Scale bar = 100 nm. (b) The particle size distribution of exosomes measured by nanoparticle tracking analysis. (c) Exosomes surface markers (HSP70, CD63, and CD81) detected using western blot.
Figure Legend Snippet: Characterization of the exosomes isolated from the peritoneal dialysis effluent (PDE). (a) Morphology of the exosomes observed by a transmission electron microscopy. Scale bar = 100 nm. (b) The particle size distribution of exosomes measured by nanoparticle tracking analysis. (c) Exosomes surface markers (HSP70, CD63, and CD81) detected using western blot.

Techniques Used: Isolation, Transmission Assay, Electron Microscopy, Western Blot

2) Product Images from "Exosomes secreted by adipose-derived mesenchymal stem cells regulate type I collagen metabolism in fibroblasts from women with stress urinary incontinence"

Article Title: Exosomes secreted by adipose-derived mesenchymal stem cells regulate type I collagen metabolism in fibroblasts from women with stress urinary incontinence

Journal: Stem Cell Research & Therapy

doi: 10.1186/s13287-018-0899-9

Isolation and characterization of adMSC-Exos. a Electron micrograph of exosomes isolated from adMSC-conditioned medium. The arrowheads indicate exosomes. Original scale bars = 0.5 μm. b Western blotting was performed with adMSC-Exos ( Exos ) or adMSC-conditioned medium ( Medium ). CD63, HSP70 and CD81 expression levels in exosomes were detected. c Fibroblasts were incubated with Dil-labeled exosomes ( Dil-Exos ; Dil is shown in red ) or carrier control ( CON ), and nuclei were stained with Hoechst 33,342 ( blue ). Original scale bars = 50 μm
Figure Legend Snippet: Isolation and characterization of adMSC-Exos. a Electron micrograph of exosomes isolated from adMSC-conditioned medium. The arrowheads indicate exosomes. Original scale bars = 0.5 μm. b Western blotting was performed with adMSC-Exos ( Exos ) or adMSC-conditioned medium ( Medium ). CD63, HSP70 and CD81 expression levels in exosomes were detected. c Fibroblasts were incubated with Dil-labeled exosomes ( Dil-Exos ; Dil is shown in red ) or carrier control ( CON ), and nuclei were stained with Hoechst 33,342 ( blue ). Original scale bars = 50 μm

Techniques Used: Isolation, Western Blot, Expressing, Incubation, Labeling, Staining

3) Product Images from "Zhen-Wu-Tang Protects IgA Nephropathy in Rats by Regulating Exosomes to Inhibit NF-κB/NLRP3 Pathway"

Article Title: Zhen-Wu-Tang Protects IgA Nephropathy in Rats by Regulating Exosomes to Inhibit NF-κB/NLRP3 Pathway

Journal: Frontiers in Pharmacology

doi: 10.3389/fphar.2020.01080

GW4869 had a negative effect on the release of exosomes. (A) Exosomal protein levels in the supernatant of HK-2 cells after GW4869 treatment were analyzed by western blot, 40 μM was the optimal concentration to inhibit the expressions of CD9 and CD81 (n = 3). The statistical data of all the proteins were analyzed and transformed by ImageJ 1.8.0 112 software. Data were expressed as mean ± SD. # P
Figure Legend Snippet: GW4869 had a negative effect on the release of exosomes. (A) Exosomal protein levels in the supernatant of HK-2 cells after GW4869 treatment were analyzed by western blot, 40 μM was the optimal concentration to inhibit the expressions of CD9 and CD81 (n = 3). The statistical data of all the proteins were analyzed and transformed by ImageJ 1.8.0 112 software. Data were expressed as mean ± SD. # P

Techniques Used: Western Blot, Concentration Assay, Transformation Assay, Software

Zhen-wu-tang (ZWT) was able to reinforce the secretion of exosomes in immunoglobulin A nephropathy (IgAN) rats. (A) CD63 expression in the glomerulus was evaluated by confocal imaging at ×200 magnification. Scale bar: 20 μm. (B, C) Western blotting of CD9, CD63, and CD81 expressed in the glomerulus of IgAN rats. (n= 3).All values were presented as mean ± SD. # P
Figure Legend Snippet: Zhen-wu-tang (ZWT) was able to reinforce the secretion of exosomes in immunoglobulin A nephropathy (IgAN) rats. (A) CD63 expression in the glomerulus was evaluated by confocal imaging at ×200 magnification. Scale bar: 20 μm. (B, C) Western blotting of CD9, CD63, and CD81 expressed in the glomerulus of IgAN rats. (n= 3).All values were presented as mean ± SD. # P

Techniques Used: Expressing, Imaging, Western Blot

The characterization of exosomes and intervention of Zhen-wu-tang (ZWT). HK-2 cells were subjected to lipopolysaccharide (LPS) (100 ng/ml) and treated with or without ZWTS (2.5%, 5%, 10%, 15%, and 20%) for 24 h. (A) Cell vitality of HK-2 cells was examined by MTT assay. (n = 6). (B) Exosomes derived from HK-2 cells were identified by transmission electron microscopy (TEM). Magnification: ×15,000. Scale bar: 200 nm. (n = 3). (C) Size distribution of exosomes extracted from HK-2 cells conditioned medium by nanoparticle analysis (NTA). (n = 3). (D) Exosomal marker proteins including CD9, CD81, and CD63 in the supernatant of HK-2 cells were determined by western blot. (n = 3). The statistical data of all the proteins were analyzed and transformed by ImageJ 1.8.0 112 software. All values were shown as mean ± SD. # P
Figure Legend Snippet: The characterization of exosomes and intervention of Zhen-wu-tang (ZWT). HK-2 cells were subjected to lipopolysaccharide (LPS) (100 ng/ml) and treated with or without ZWTS (2.5%, 5%, 10%, 15%, and 20%) for 24 h. (A) Cell vitality of HK-2 cells was examined by MTT assay. (n = 6). (B) Exosomes derived from HK-2 cells were identified by transmission electron microscopy (TEM). Magnification: ×15,000. Scale bar: 200 nm. (n = 3). (C) Size distribution of exosomes extracted from HK-2 cells conditioned medium by nanoparticle analysis (NTA). (n = 3). (D) Exosomal marker proteins including CD9, CD81, and CD63 in the supernatant of HK-2 cells were determined by western blot. (n = 3). The statistical data of all the proteins were analyzed and transformed by ImageJ 1.8.0 112 software. All values were shown as mean ± SD. # P

Techniques Used: MTT Assay, Derivative Assay, Transmission Assay, Electron Microscopy, Transmission Electron Microscopy, Marker, Western Blot, Transformation Assay, Software

4) Product Images from "The Ubiquitin-Specific Protease 18 Promotes Hepatitis C Virus Production by Increasing Viral Infectivity"

Article Title: The Ubiquitin-Specific Protease 18 Promotes Hepatitis C Virus Production by Increasing Viral Infectivity

Journal: Mediators of Inflammation

doi: 10.1155/2019/3124745

USP18 promotes HCV infectivity and CD81 expression in Huh7.5 cells. (a) Huh7.5 cells were seeded at 3 × 10 5 /ml, 2 ml per well in 6-well plates in antibiotic-free medium for 24 hours before 1 μ g, 2 μ g, 4 μ g, and 6 μ g pcDNA3.1-USP18 or 4 μ g empty vector pcDNA3.1 was transfected into each well. 48 hours posttransfection, total RNA was extracted; USP18 and CD81 expression was quantified by qRT-PCR and normalized to GAPDH expression as previously described. (b) Huh7.5 cells were seeded at 3 × 10 5 /ml, 2 ml per well in 6-well plates in antibiotic-free medium for 24 hours before 4 μ g pcDNA3.1-USP18 was transfected into each well. (B-1) Lower-left and lower right quadrants indicate that the cells were transfected with empty vector pcDNA3.1 or pcDNA3.1-USP18 at 72 hours, with the percentage of cells in respective quadrants. All dot blots show CD81 expression (vertical axis) according to USP18 expression (horizontal axis). Upper-left and right quadrants show the range of isotype control for PE- and Alexa 488-conjugated antibodies. (B-2) Huh7.5 cells with transfected empty vector pcDNA3.1 (open histogram, dark green line) or pcDNA3.1-USP18 DNA (filled histogram) were stained with a mouse anti-flag primary/FITC-conjugated goat anti-mouse secondary antibody, rabbit anti-human USP18 primary/Alexa 488-conjugated goat anti-rabbit secondary antibody, and PE-conjugated anti-human CD81 monoclonal antibody. The cells transfected with pcDNA3.1-USP18 were also stained with mouse and rabbit IgG and isotype control antibodies (open histogram, brown line). (B-3) Comparison of USP18/CD81 expression in or on Huh7.5 cells with sham control, transfected pcDNA3.1 control, and pcDNA3.1-USP18 DNA. Bars indicate MFI of USP18 (FITC and Alexa 488)/CD81 (PE) expression ± STDV. Untreated: untreated control; pcDNA3.1: transfected with 4 μ g empty vector pcDNA3.1; 1 μ g, 2 μ g, 4 μ g, and 6 μ g: transfected with 1 μ g, 2 μ g, 4 μ g, or 6 μ g pcDNA3.1-USP18; sham control: untreated Huh7.5 cells only. Results are presented as means ± SD ( n ≥ 3). ∗ p
Figure Legend Snippet: USP18 promotes HCV infectivity and CD81 expression in Huh7.5 cells. (a) Huh7.5 cells were seeded at 3 × 10 5 /ml, 2 ml per well in 6-well plates in antibiotic-free medium for 24 hours before 1 μ g, 2 μ g, 4 μ g, and 6 μ g pcDNA3.1-USP18 or 4 μ g empty vector pcDNA3.1 was transfected into each well. 48 hours posttransfection, total RNA was extracted; USP18 and CD81 expression was quantified by qRT-PCR and normalized to GAPDH expression as previously described. (b) Huh7.5 cells were seeded at 3 × 10 5 /ml, 2 ml per well in 6-well plates in antibiotic-free medium for 24 hours before 4 μ g pcDNA3.1-USP18 was transfected into each well. (B-1) Lower-left and lower right quadrants indicate that the cells were transfected with empty vector pcDNA3.1 or pcDNA3.1-USP18 at 72 hours, with the percentage of cells in respective quadrants. All dot blots show CD81 expression (vertical axis) according to USP18 expression (horizontal axis). Upper-left and right quadrants show the range of isotype control for PE- and Alexa 488-conjugated antibodies. (B-2) Huh7.5 cells with transfected empty vector pcDNA3.1 (open histogram, dark green line) or pcDNA3.1-USP18 DNA (filled histogram) were stained with a mouse anti-flag primary/FITC-conjugated goat anti-mouse secondary antibody, rabbit anti-human USP18 primary/Alexa 488-conjugated goat anti-rabbit secondary antibody, and PE-conjugated anti-human CD81 monoclonal antibody. The cells transfected with pcDNA3.1-USP18 were also stained with mouse and rabbit IgG and isotype control antibodies (open histogram, brown line). (B-3) Comparison of USP18/CD81 expression in or on Huh7.5 cells with sham control, transfected pcDNA3.1 control, and pcDNA3.1-USP18 DNA. Bars indicate MFI of USP18 (FITC and Alexa 488)/CD81 (PE) expression ± STDV. Untreated: untreated control; pcDNA3.1: transfected with 4 μ g empty vector pcDNA3.1; 1 μ g, 2 μ g, 4 μ g, and 6 μ g: transfected with 1 μ g, 2 μ g, 4 μ g, or 6 μ g pcDNA3.1-USP18; sham control: untreated Huh7.5 cells only. Results are presented as means ± SD ( n ≥ 3). ∗ p

Techniques Used: Infection, Expressing, Plasmid Preparation, Transfection, Quantitative RT-PCR, Staining

5) Product Images from "Exosomes secreted by adipose-derived mesenchymal stem cells regulate type I collagen metabolism in fibroblasts from women with stress urinary incontinence"

Article Title: Exosomes secreted by adipose-derived mesenchymal stem cells regulate type I collagen metabolism in fibroblasts from women with stress urinary incontinence

Journal: Stem Cell Research & Therapy

doi: 10.1186/s13287-018-0899-9

Isolation and characterization of adMSC-Exos. a Electron micrograph of exosomes isolated from adMSC-conditioned medium. The arrowheads indicate exosomes. Original scale bars = 0.5 μm. b Western blotting was performed with adMSC-Exos ( Exos ) or adMSC-conditioned medium ( Medium ). CD63, HSP70 and CD81 expression levels in exosomes were detected. c Fibroblasts were incubated with Dil-labeled exosomes ( Dil-Exos ; Dil is shown in red ) or carrier control ( CON ), and nuclei were stained with Hoechst 33,342 ( blue ). Original scale bars = 50 μm
Figure Legend Snippet: Isolation and characterization of adMSC-Exos. a Electron micrograph of exosomes isolated from adMSC-conditioned medium. The arrowheads indicate exosomes. Original scale bars = 0.5 μm. b Western blotting was performed with adMSC-Exos ( Exos ) or adMSC-conditioned medium ( Medium ). CD63, HSP70 and CD81 expression levels in exosomes were detected. c Fibroblasts were incubated with Dil-labeled exosomes ( Dil-Exos ; Dil is shown in red ) or carrier control ( CON ), and nuclei were stained with Hoechst 33,342 ( blue ). Original scale bars = 50 μm

Techniques Used: Isolation, Western Blot, Expressing, Incubation, Labeling, Staining

6) Product Images from "Stable expression of a truncated TLX variant drives differentiation of induced pluripotent stem cells into self-renewing neural stem cells for production of extracellular vesicles"

Article Title: Stable expression of a truncated TLX variant drives differentiation of induced pluripotent stem cells into self-renewing neural stem cells for production of extracellular vesicles

Journal: Stem Cell Research & Therapy

doi: 10.1186/s13287-022-03131-4

Purification and characterization of iNSC-EVs. A Scheme of purification steps of iNSC-EVs. B Nanoflow cytometry (nanoFCM) results of iNSC-EVs (blue), iNSC-EVs treated with Triton X-100 (red) and Triton (green) after the TFF step, fractions of DGUC (7–11) and QA ( D ). C Concentration of DGUC fraction detected by nanoFCM. TEM results of iNSC-EVs after the TFF step ( E ), QA ( G ) and a fraction (1–6) or fraction (7–11) of DGUC ( H ). I Immunoblotting of iNSCs TLX−TP or purified iNSC-EVs for Flag, CD63 and CD81 expressions
Figure Legend Snippet: Purification and characterization of iNSC-EVs. A Scheme of purification steps of iNSC-EVs. B Nanoflow cytometry (nanoFCM) results of iNSC-EVs (blue), iNSC-EVs treated with Triton X-100 (red) and Triton (green) after the TFF step, fractions of DGUC (7–11) and QA ( D ). C Concentration of DGUC fraction detected by nanoFCM. TEM results of iNSC-EVs after the TFF step ( E ), QA ( G ) and a fraction (1–6) or fraction (7–11) of DGUC ( H ). I Immunoblotting of iNSCs TLX−TP or purified iNSC-EVs for Flag, CD63 and CD81 expressions

Techniques Used: Purification, Cytometry, Concentration Assay, Transmission Electron Microscopy

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    Abcam anti cd8 antibody
    CD4 + TILs colocalize with PD-L1 + cells and contribute to poor survival. (a) Representative multi-IF images showing a sample with more CD4 + TILs than <t>CD8</t> + TILs in the stroma. Scale bar, 100 μ m. (b) Representative multi-IF images showing a sample with more CD8 + TILs than CD4 + TILs in the stroma. Scale bar, 100 μ m. (c) A pie chart was plotted according to IHC staining results showing 84% of patients have more CD4 + TILs than CD8 + TILs in the stroma. (d) Representative multi-IF images showing a sample with PD-L1 + cells colocalizing more with CD4 + TILs than with CD8 + TILs in the stroma. Scale bar, 100 μ m. (e) The Kaplan–Meier survival analysis showing patients with elevated levels of CD4 + TILs (red line, n = 26) have poor OS, compared to patients with low levels of CD4 + TILs (blue line, n = 51). (f) The Kaplan–Meier survival analysis showing patients with a low CD8/CD4 ratio (blue line, n = 46) have poor OS, compared to patients with a high CD8/CD4 ratio (red line, n = 31).
    Anti Cd8 Antibody, supplied by Abcam, used in various techniques. Bioz Stars score: 88/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti cd8 antibody/product/Abcam
    Average 88 stars, based on 1 article reviews
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    cd81  (Abcam)
    99
    Abcam cd81
    Gene expression analysis of CD9 and <t>CD81</t> in prostate cancer compared to normal samples in publicly available gene expression profiling datasets. (A–D): Gene expression boxplots for CD9. (A) TCGA dataset-prostate adenocarcinoma versus normal. (B) Wallace dataset- prostate adenocarcinoma vs. normal. (C) Lapointe dataset- prostate carcinoma vs. normal. (D) Yu dataset-prostate carcinoma vs. normal. ( E -H): Gene expression boxplots for CD81. (E) TCGA dataset-prostate adenocarcinoma vs. normal. (F) Vanaja dataset-prostate adenocarcinoma vs. normal. (G) Taylor dataset-prostate carcinoma vs. normal. (H) Singh dataset- prostate carcinoma vs. normal. 1, Normal; 2, Carcinoma or Adenocarcinoma.
    Cd81, supplied by Abcam, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Abcam rat anti mouse cd8 monoclonal antibody
    A : Fluorescence microscopy revealed lower protein expression of B2M in MN-siB2M–treated grafts compared with control probe grafts 2 weeks after adoptive transfer (green, insulin; red, B2M; blue, DAPI nuclear stain; magnification bar = 50 μm). B : Fluorescence microscopy showed markedly lower <t>CD8</t> + T-cell infiltration in MN-siB2M–treated grafts compared with control islet grafts 2 weeks after adoptive transfer (green, insulin; red, CD8; blue, DAPI nuclear stain; magnification bar = 40 μm). C : Western blot analysis confirmed notably lower B2M protein expression in MN-siB2M–treated islet grafts compared with control grafts. (A high-quality digital representation of this figure is available in the online issue.)
    Rat Anti Mouse Cd8 Monoclonal Antibody, supplied by Abcam, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Abcam mouse anti cd81
    Immunolocalization of exosome markers in the RPE/choroid complex. (A): CD63 (red); AMD eye from a 74-year-old male. Anti-CD63 antibody labels large amorphous areas (CD63: arrows) in drusen. <t>(B):CD81</t> (red); AMD eye from a 96-year-old male (CD81, arrows). (C): LAMP2 (red). AMD eye from a 93-year-old (LAMP2, arrow). (D): CD63 (red); Non-AMD eye from a 75-year-old male as an age-matched control. No CD63 labeling was seen in the drusen from any age-matched control (n = 10). (E–H): CD63 (green) co-localization with proteins that are known to be in drusen. (E) amyloid β (red) showed no co-localization with CD63 (arrow). (F) α B crystalline (red) showed co-localization with CD63 (arrowhead). (G) C5b-9 (red) showed no co-localization with CD63 (arrow). (H) CFH (red) showed co-localization with CD63 (arrowhead). Dr, drusen. Blue: DAPI.
    Mouse Anti Cd81, supplied by Abcam, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    CD4 + TILs colocalize with PD-L1 + cells and contribute to poor survival. (a) Representative multi-IF images showing a sample with more CD4 + TILs than CD8 + TILs in the stroma. Scale bar, 100 μ m. (b) Representative multi-IF images showing a sample with more CD8 + TILs than CD4 + TILs in the stroma. Scale bar, 100 μ m. (c) A pie chart was plotted according to IHC staining results showing 84% of patients have more CD4 + TILs than CD8 + TILs in the stroma. (d) Representative multi-IF images showing a sample with PD-L1 + cells colocalizing more with CD4 + TILs than with CD8 + TILs in the stroma. Scale bar, 100 μ m. (e) The Kaplan–Meier survival analysis showing patients with elevated levels of CD4 + TILs (red line, n = 26) have poor OS, compared to patients with low levels of CD4 + TILs (blue line, n = 51). (f) The Kaplan–Meier survival analysis showing patients with a low CD8/CD4 ratio (blue line, n = 46) have poor OS, compared to patients with a high CD8/CD4 ratio (red line, n = 31).

    Journal: Journal of Immunology Research

    Article Title: Tumor-Infiltrating Immune Cells and PD-L1 as Prognostic Biomarkers in Primary Esophageal Small Cell Carcinoma

    doi: 10.1155/2020/8884683

    Figure Lengend Snippet: CD4 + TILs colocalize with PD-L1 + cells and contribute to poor survival. (a) Representative multi-IF images showing a sample with more CD4 + TILs than CD8 + TILs in the stroma. Scale bar, 100 μ m. (b) Representative multi-IF images showing a sample with more CD8 + TILs than CD4 + TILs in the stroma. Scale bar, 100 μ m. (c) A pie chart was plotted according to IHC staining results showing 84% of patients have more CD4 + TILs than CD8 + TILs in the stroma. (d) Representative multi-IF images showing a sample with PD-L1 + cells colocalizing more with CD4 + TILs than with CD8 + TILs in the stroma. Scale bar, 100 μ m. (e) The Kaplan–Meier survival analysis showing patients with elevated levels of CD4 + TILs (red line, n = 26) have poor OS, compared to patients with low levels of CD4 + TILs (blue line, n = 51). (f) The Kaplan–Meier survival analysis showing patients with a low CD8/CD4 ratio (blue line, n = 46) have poor OS, compared to patients with a high CD8/CD4 ratio (red line, n = 31).

    Article Snippet: Immunohistochemistry StainingIHC staining was performed using anti-PD-L1 antibody (1 : 500) (clone 28-8, Abcam, Cambridge, UK) [ – ], anti-CD4 antibody (1 : 500) (clone EPR6855, Abcam, Cambridge, UK), anti-CD8 antibody (1 : 200) (rabbit polyclonal anti-CD8, Abcam, Cambridge, UK), anti-CD163 antibody (1 : 500) (clone EPR19518, Abcam, Cambridge, UK), and anti-FoxP3 antibody (1 : 100) (clone 236A/E7, Abcam, Cambridge, UK) as the primary antibodies.

    Techniques: Immunohistochemistry, Staining

    Expression and prognostic value of FoxP3 in PESCC. (a) Representative multi-IF images of two samples showing FoxP3/CD4 ratios in the stroma. Scale bar, 100 μ m. (b) Representative multi-IF images showing two samples with high (left) and low (right) FoxP3/CD8 ratios in stroma. Scale bar, 100 μ m. (c) Rich FoxP3 (red line, n = 26) was associated with significantly shorter OS than poor FoxP3 (blue line, n = 51). (d) A high FoxP3/CD8 ratio (red line, n = 28) was associated with shorter OS than a low FoxP3/CD8 ratio (blue line, n = 49).

    Journal: Journal of Immunology Research

    Article Title: Tumor-Infiltrating Immune Cells and PD-L1 as Prognostic Biomarkers in Primary Esophageal Small Cell Carcinoma

    doi: 10.1155/2020/8884683

    Figure Lengend Snippet: Expression and prognostic value of FoxP3 in PESCC. (a) Representative multi-IF images of two samples showing FoxP3/CD4 ratios in the stroma. Scale bar, 100 μ m. (b) Representative multi-IF images showing two samples with high (left) and low (right) FoxP3/CD8 ratios in stroma. Scale bar, 100 μ m. (c) Rich FoxP3 (red line, n = 26) was associated with significantly shorter OS than poor FoxP3 (blue line, n = 51). (d) A high FoxP3/CD8 ratio (red line, n = 28) was associated with shorter OS than a low FoxP3/CD8 ratio (blue line, n = 49).

    Article Snippet: Immunohistochemistry StainingIHC staining was performed using anti-PD-L1 antibody (1 : 500) (clone 28-8, Abcam, Cambridge, UK) [ – ], anti-CD4 antibody (1 : 500) (clone EPR6855, Abcam, Cambridge, UK), anti-CD8 antibody (1 : 200) (rabbit polyclonal anti-CD8, Abcam, Cambridge, UK), anti-CD163 antibody (1 : 500) (clone EPR19518, Abcam, Cambridge, UK), and anti-FoxP3 antibody (1 : 100) (clone 236A/E7, Abcam, Cambridge, UK) as the primary antibodies.

    Techniques: Expressing

    The expression of PD-L1 in the stroma correlates with tumor-infiltrating immune cells in PESCC. (a) Representative immunohistochemistry (IHC) images showing different expression patterns of PD-L1 in PESCC tissues. Scale bar, 100 μ m. S: stroma; T: tumor. (b) Distribution of different expression patterns of PD-L1 are plotted in the pie chart. (c) The Kaplan–Meier survival analysis of overall survival (OS) in a cohort of 77 PESCC patients according to positive (red line, n = 26) and negative (blue line, n = 51) PD-L1 expression. (d) Representative IHC images showing PD-L1, CD4, CD8, and CD163 expression in the stroma using consecutive sections of PD-L1-positive and PD-L1-negative specimens. Scale bar, 100 μ m. S: stroma; T: tumor. (e) The presence of PD-L1 is positively associated with the number of CD4 + TILs, CD8 + TILs, and CD163 + TAMs ( ∗∗ p

    Journal: Journal of Immunology Research

    Article Title: Tumor-Infiltrating Immune Cells and PD-L1 as Prognostic Biomarkers in Primary Esophageal Small Cell Carcinoma

    doi: 10.1155/2020/8884683

    Figure Lengend Snippet: The expression of PD-L1 in the stroma correlates with tumor-infiltrating immune cells in PESCC. (a) Representative immunohistochemistry (IHC) images showing different expression patterns of PD-L1 in PESCC tissues. Scale bar, 100 μ m. S: stroma; T: tumor. (b) Distribution of different expression patterns of PD-L1 are plotted in the pie chart. (c) The Kaplan–Meier survival analysis of overall survival (OS) in a cohort of 77 PESCC patients according to positive (red line, n = 26) and negative (blue line, n = 51) PD-L1 expression. (d) Representative IHC images showing PD-L1, CD4, CD8, and CD163 expression in the stroma using consecutive sections of PD-L1-positive and PD-L1-negative specimens. Scale bar, 100 μ m. S: stroma; T: tumor. (e) The presence of PD-L1 is positively associated with the number of CD4 + TILs, CD8 + TILs, and CD163 + TAMs ( ∗∗ p

    Article Snippet: Immunohistochemistry StainingIHC staining was performed using anti-PD-L1 antibody (1 : 500) (clone 28-8, Abcam, Cambridge, UK) [ – ], anti-CD4 antibody (1 : 500) (clone EPR6855, Abcam, Cambridge, UK), anti-CD8 antibody (1 : 200) (rabbit polyclonal anti-CD8, Abcam, Cambridge, UK), anti-CD163 antibody (1 : 500) (clone EPR19518, Abcam, Cambridge, UK), and anti-FoxP3 antibody (1 : 100) (clone 236A/E7, Abcam, Cambridge, UK) as the primary antibodies.

    Techniques: Expressing, Immunohistochemistry

    CD4 + TILs colocalize with PD-L1 + CD163 + TAMs. Representative multicolor immunofluorescence (multi-IF) images showing (a) high and (b) low colocalization of PD-L1 + cells with CD163 + TAMs. Scale bar, 100 μ m. Representative multi-IF images showing costaining of (c) CD4 + PD-L1 + CD163 + TILs and (d) CD8 + PD-L1 + CD163 + TILs in serial sections of the same specimen. Cells were counterstained with DAPI (blue, nucleus). Scale bar, 100 μ m.

    Journal: Journal of Immunology Research

    Article Title: Tumor-Infiltrating Immune Cells and PD-L1 as Prognostic Biomarkers in Primary Esophageal Small Cell Carcinoma

    doi: 10.1155/2020/8884683

    Figure Lengend Snippet: CD4 + TILs colocalize with PD-L1 + CD163 + TAMs. Representative multicolor immunofluorescence (multi-IF) images showing (a) high and (b) low colocalization of PD-L1 + cells with CD163 + TAMs. Scale bar, 100 μ m. Representative multi-IF images showing costaining of (c) CD4 + PD-L1 + CD163 + TILs and (d) CD8 + PD-L1 + CD163 + TILs in serial sections of the same specimen. Cells were counterstained with DAPI (blue, nucleus). Scale bar, 100 μ m.

    Article Snippet: Immunohistochemistry StainingIHC staining was performed using anti-PD-L1 antibody (1 : 500) (clone 28-8, Abcam, Cambridge, UK) [ – ], anti-CD4 antibody (1 : 500) (clone EPR6855, Abcam, Cambridge, UK), anti-CD8 antibody (1 : 200) (rabbit polyclonal anti-CD8, Abcam, Cambridge, UK), anti-CD163 antibody (1 : 500) (clone EPR19518, Abcam, Cambridge, UK), and anti-FoxP3 antibody (1 : 100) (clone 236A/E7, Abcam, Cambridge, UK) as the primary antibodies.

    Techniques: Immunofluorescence

    Gene expression analysis of CD9 and CD81 in prostate cancer compared to normal samples in publicly available gene expression profiling datasets. (A–D): Gene expression boxplots for CD9. (A) TCGA dataset-prostate adenocarcinoma versus normal. (B) Wallace dataset- prostate adenocarcinoma vs. normal. (C) Lapointe dataset- prostate carcinoma vs. normal. (D) Yu dataset-prostate carcinoma vs. normal. ( E -H): Gene expression boxplots for CD81. (E) TCGA dataset-prostate adenocarcinoma vs. normal. (F) Vanaja dataset-prostate adenocarcinoma vs. normal. (G) Taylor dataset-prostate carcinoma vs. normal. (H) Singh dataset- prostate carcinoma vs. normal. 1, Normal; 2, Carcinoma or Adenocarcinoma.

    Journal: Matrix biology : journal of the International Society for Matrix Biology

    Article Title: Prostate cancer sheds the αvβ3 integrin in vivo through exosomes

    doi: 10.1016/j.matbio.2018.08.004

    Figure Lengend Snippet: Gene expression analysis of CD9 and CD81 in prostate cancer compared to normal samples in publicly available gene expression profiling datasets. (A–D): Gene expression boxplots for CD9. (A) TCGA dataset-prostate adenocarcinoma versus normal. (B) Wallace dataset- prostate adenocarcinoma vs. normal. (C) Lapointe dataset- prostate carcinoma vs. normal. (D) Yu dataset-prostate carcinoma vs. normal. ( E -H): Gene expression boxplots for CD81. (E) TCGA dataset-prostate adenocarcinoma vs. normal. (F) Vanaja dataset-prostate adenocarcinoma vs. normal. (G) Taylor dataset-prostate carcinoma vs. normal. (H) Singh dataset- prostate carcinoma vs. normal. 1, Normal; 2, Carcinoma or Adenocarcinoma.

    Article Snippet: The following antibodies (Abs) were used for immunoblotting (IB) analysis: mouse monoclonal Abs to PSMA (ab-19,071; Abcam), CD9 (sc-13,118; Santa Cruz Biotechnology), CD81 (ab-23,505; Abcam), CD63 (ab-8219; Abcam), GM130 (61,820; BD Biosciences), EpCAM (2929; Cell Signaling Technology), TUBULIN (T-8535; Sigma); rabbit mAb to TSG101 (ab-125,011; Abcam); rabbit polyclonal Abs (pAbs) to FLOTILLIN1 (FLOT1) (ab-41,927; Abcam), GFP (ab-6556; Abcam), CALNEXIN (CANX) (sc-11,397; Santa Cruz Biotechnology), ERK1 (sc-93; Santa Cruz Biotechnology), and Rb-IgG (Sigma).

    Techniques: Expressing

    GFP-tagged αvβ3 integrin is expressed in prostate cancer cell ExVs, and it is localized in distant lesions in vivo. (A) IB analysis of expression of GFP in total cell lysates (TCL) from parental C4-2B, C4-2B-GFP (Mock) and C4-2B-β3-GFP cells. Calnexin (CANX) was used as loading control. (B) IB analysis of expression of αv, β3, CD63, CD81 (non-reducing conditions) and GFP, β3, TSG101, CD9 (reducing conditions) in lysates from ExVs derived from C4-2B-β3-GFP cells by differential ultracentrifugation. (C) In vivo fluorescence and corresponding phase contrast images are shown for liver, DU145 tumor, and prostate isolated from NSG mice that were injected subcutaneously with C4-2B-β3-GFP cells on the right side and non-transfected DU145 cells on the left side. Two representative samples are shown. (D) NTA for size distribution and concentration of GFP-positive ExVs isolated by differential ultracentrifugation from plasma of the mice subcutaneously injected with C4-2B-β3-GFP cells. One representative sample is shown.

    Journal: Matrix biology : journal of the International Society for Matrix Biology

    Article Title: Prostate cancer sheds the αvβ3 integrin in vivo through exosomes

    doi: 10.1016/j.matbio.2018.08.004

    Figure Lengend Snippet: GFP-tagged αvβ3 integrin is expressed in prostate cancer cell ExVs, and it is localized in distant lesions in vivo. (A) IB analysis of expression of GFP in total cell lysates (TCL) from parental C4-2B, C4-2B-GFP (Mock) and C4-2B-β3-GFP cells. Calnexin (CANX) was used as loading control. (B) IB analysis of expression of αv, β3, CD63, CD81 (non-reducing conditions) and GFP, β3, TSG101, CD9 (reducing conditions) in lysates from ExVs derived from C4-2B-β3-GFP cells by differential ultracentrifugation. (C) In vivo fluorescence and corresponding phase contrast images are shown for liver, DU145 tumor, and prostate isolated from NSG mice that were injected subcutaneously with C4-2B-β3-GFP cells on the right side and non-transfected DU145 cells on the left side. Two representative samples are shown. (D) NTA for size distribution and concentration of GFP-positive ExVs isolated by differential ultracentrifugation from plasma of the mice subcutaneously injected with C4-2B-β3-GFP cells. One representative sample is shown.

    Article Snippet: The following antibodies (Abs) were used for immunoblotting (IB) analysis: mouse monoclonal Abs to PSMA (ab-19,071; Abcam), CD9 (sc-13,118; Santa Cruz Biotechnology), CD81 (ab-23,505; Abcam), CD63 (ab-8219; Abcam), GM130 (61,820; BD Biosciences), EpCAM (2929; Cell Signaling Technology), TUBULIN (T-8535; Sigma); rabbit mAb to TSG101 (ab-125,011; Abcam); rabbit polyclonal Abs (pAbs) to FLOTILLIN1 (FLOT1) (ab-41,927; Abcam), GFP (ab-6556; Abcam), CALNEXIN (CANX) (sc-11,397; Santa Cruz Biotechnology), ERK1 (sc-93; Santa Cruz Biotechnology), and Rb-IgG (Sigma).

    Techniques: In Vivo, Expressing, Derivative Assay, Fluorescence, Isolation, Mouse Assay, Injection, Transfection, Concentration Assay

    Size distribution analysis and differences in β3 integrin levels in ExVs from plasma of prostate cancer patients compared to subjects not affected by cancer. (A) NTA of ExVs isolated by differential ultracentrifugation from plasma of individuals not affected by cancer (left panel) and prostate cancer patients (right panel). (B) IB analysis of β3, CD9, CD81, and αv levels in ExVs isolated by differential ultracentrifugation from plasma of prostate cancer patients compared to age-matched individuals not affected by cancer. CANX was analyzed as a marker absent in exosomes. Lanes 1, 2, and 3: EV lysates isolated after the plasma was pooled from at least two subjects not affected by cancer (total of 7 biological samples represented in 3 lanes); lanes 4–8: EV lysates from individual patients. 30 μg of exosome lysates were loaded in each lane. (C) lodixanol gradient purified Exosomes (Exo) from prostate cancer patient plasma (pooled from n = 3) were immunocaptured with an antibody to Prostate Specific Membrane Antigen (PSMA) or isotype rabbit immunoglobulin (Rb-IgG) conjugated with Dynabeads M-270 epoxy magnetic beads, according to the manufacturer’s protocol. The immunocaptured whole exosomes were lysed with RIPA buffer, and lysates were separated by SDS-PAGE (7.5% gel). IB analysis shows expression of β3, CD9 and CD63 (exosomal markers), Trop-2, and PSMA; in contrast, TSG101 (exosomal marker), CANX and EpCAM were not detected. HC-IgG, heavy chain IgG.

    Journal: Matrix biology : journal of the International Society for Matrix Biology

    Article Title: Prostate cancer sheds the αvβ3 integrin in vivo through exosomes

    doi: 10.1016/j.matbio.2018.08.004

    Figure Lengend Snippet: Size distribution analysis and differences in β3 integrin levels in ExVs from plasma of prostate cancer patients compared to subjects not affected by cancer. (A) NTA of ExVs isolated by differential ultracentrifugation from plasma of individuals not affected by cancer (left panel) and prostate cancer patients (right panel). (B) IB analysis of β3, CD9, CD81, and αv levels in ExVs isolated by differential ultracentrifugation from plasma of prostate cancer patients compared to age-matched individuals not affected by cancer. CANX was analyzed as a marker absent in exosomes. Lanes 1, 2, and 3: EV lysates isolated after the plasma was pooled from at least two subjects not affected by cancer (total of 7 biological samples represented in 3 lanes); lanes 4–8: EV lysates from individual patients. 30 μg of exosome lysates were loaded in each lane. (C) lodixanol gradient purified Exosomes (Exo) from prostate cancer patient plasma (pooled from n = 3) were immunocaptured with an antibody to Prostate Specific Membrane Antigen (PSMA) or isotype rabbit immunoglobulin (Rb-IgG) conjugated with Dynabeads M-270 epoxy magnetic beads, according to the manufacturer’s protocol. The immunocaptured whole exosomes were lysed with RIPA buffer, and lysates were separated by SDS-PAGE (7.5% gel). IB analysis shows expression of β3, CD9 and CD63 (exosomal markers), Trop-2, and PSMA; in contrast, TSG101 (exosomal marker), CANX and EpCAM were not detected. HC-IgG, heavy chain IgG.

    Article Snippet: The following antibodies (Abs) were used for immunoblotting (IB) analysis: mouse monoclonal Abs to PSMA (ab-19,071; Abcam), CD9 (sc-13,118; Santa Cruz Biotechnology), CD81 (ab-23,505; Abcam), CD63 (ab-8219; Abcam), GM130 (61,820; BD Biosciences), EpCAM (2929; Cell Signaling Technology), TUBULIN (T-8535; Sigma); rabbit mAb to TSG101 (ab-125,011; Abcam); rabbit polyclonal Abs (pAbs) to FLOTILLIN1 (FLOT1) (ab-41,927; Abcam), GFP (ab-6556; Abcam), CALNEXIN (CANX) (sc-11,397; Santa Cruz Biotechnology), ERK1 (sc-93; Santa Cruz Biotechnology), and Rb-IgG (Sigma).

    Techniques: Isolation, Marker, Purification, Magnetic Beads, SDS Page, Expressing

    A : Fluorescence microscopy revealed lower protein expression of B2M in MN-siB2M–treated grafts compared with control probe grafts 2 weeks after adoptive transfer (green, insulin; red, B2M; blue, DAPI nuclear stain; magnification bar = 50 μm). B : Fluorescence microscopy showed markedly lower CD8 + T-cell infiltration in MN-siB2M–treated grafts compared with control islet grafts 2 weeks after adoptive transfer (green, insulin; red, CD8; blue, DAPI nuclear stain; magnification bar = 40 μm). C : Western blot analysis confirmed notably lower B2M protein expression in MN-siB2M–treated islet grafts compared with control grafts. (A high-quality digital representation of this figure is available in the online issue.)

    Journal: Diabetes

    Article Title: A Theranostic Small Interfering RNA Nanoprobe Protects Pancreatic Islet Grafts From Adoptively Transferred Immune Rejection

    doi: 10.2337/db12-0441

    Figure Lengend Snippet: A : Fluorescence microscopy revealed lower protein expression of B2M in MN-siB2M–treated grafts compared with control probe grafts 2 weeks after adoptive transfer (green, insulin; red, B2M; blue, DAPI nuclear stain; magnification bar = 50 μm). B : Fluorescence microscopy showed markedly lower CD8 + T-cell infiltration in MN-siB2M–treated grafts compared with control islet grafts 2 weeks after adoptive transfer (green, insulin; red, CD8; blue, DAPI nuclear stain; magnification bar = 40 μm). C : Western blot analysis confirmed notably lower B2M protein expression in MN-siB2M–treated islet grafts compared with control grafts. (A high-quality digital representation of this figure is available in the online issue.)

    Article Snippet: For human insulin and mouse CD8+ T cells double-staining, sections were incubated with a guinea pig anti-human insulin primary antibody (1:200 dilution, Abcam) and rat anti-mouse CD8 monoclonal antibody (1:50 dilution, Abcam), followed by an FITC-labeled goat anti-guinea pig secondary IgG (1:100 dilution, Abcam) and Alexa Fluor 594 conjugated secondary goat anti-rat IgG (1:50 dilution, Invitrogen).

    Techniques: Fluorescence, Microscopy, Expressing, Adoptive Transfer Assay, Staining, Western Blot

    Immunolocalization of exosome markers in the RPE/choroid complex. (A): CD63 (red); AMD eye from a 74-year-old male. Anti-CD63 antibody labels large amorphous areas (CD63: arrows) in drusen. (B):CD81 (red); AMD eye from a 96-year-old male (CD81, arrows). (C): LAMP2 (red). AMD eye from a 93-year-old (LAMP2, arrow). (D): CD63 (red); Non-AMD eye from a 75-year-old male as an age-matched control. No CD63 labeling was seen in the drusen from any age-matched control (n = 10). (E–H): CD63 (green) co-localization with proteins that are known to be in drusen. (E) amyloid β (red) showed no co-localization with CD63 (arrow). (F) α B crystalline (red) showed co-localization with CD63 (arrowhead). (G) C5b-9 (red) showed no co-localization with CD63 (arrow). (H) CFH (red) showed co-localization with CD63 (arrowhead). Dr, drusen. Blue: DAPI.

    Journal: PLoS ONE

    Article Title: Autophagy and Exosomes in the Aged Retinal Pigment Epithelium: Possible Relevance to Drusen Formation and Age-Related Macular Degeneration

    doi: 10.1371/journal.pone.0004160

    Figure Lengend Snippet: Immunolocalization of exosome markers in the RPE/choroid complex. (A): CD63 (red); AMD eye from a 74-year-old male. Anti-CD63 antibody labels large amorphous areas (CD63: arrows) in drusen. (B):CD81 (red); AMD eye from a 96-year-old male (CD81, arrows). (C): LAMP2 (red). AMD eye from a 93-year-old (LAMP2, arrow). (D): CD63 (red); Non-AMD eye from a 75-year-old male as an age-matched control. No CD63 labeling was seen in the drusen from any age-matched control (n = 10). (E–H): CD63 (green) co-localization with proteins that are known to be in drusen. (E) amyloid β (red) showed no co-localization with CD63 (arrow). (F) α B crystalline (red) showed co-localization with CD63 (arrowhead). (G) C5b-9 (red) showed no co-localization with CD63 (arrow). (H) CFH (red) showed co-localization with CD63 (arrowhead). Dr, drusen. Blue: DAPI.

    Article Snippet: 110 µl of 1 M glycine was added (i.e., 100 mM final), mixed gently and let stand on the bench at room temperature for 30 min. Beads were re-suspended in 0.5 ml PBS/0.5% BSA, 10 µl coated beads were incubated with 50 µl primary antibodies: mouse anti-CFH (1∶50, Serotec), mouse anti-CD63 (1∶50, Abcam), mouse ant-LAMP2 (1∶50, Abcam), mouse anti-CD81 (1∶50, Abcam), mouse anti-C3 (1∶50, Abcam).

    Techniques: Labeling

    Analyses of exosomes by FACS. (A) Exoxomes are CD63, LAMP2, CD81 and C3 positive, but CFH negative. Isotype control: Black; CFH: light green; CD63: blue; LAMP2: red; CD81: dark green; C3: magenta. (B) Added CFH bound to exosomes in a dose-dependent manner. (C) Added CFH did not alter the amount of C3 on exosomes. For B and C: CFH 0.5 mg/mL: black; CFH 0.05 mg/mL: blue; CFH 0.005 mg/mL: purple; CFH 0.0005 mg/mL: green; CFH 0 mg/mL: red.

    Journal: PLoS ONE

    Article Title: Autophagy and Exosomes in the Aged Retinal Pigment Epithelium: Possible Relevance to Drusen Formation and Age-Related Macular Degeneration

    doi: 10.1371/journal.pone.0004160

    Figure Lengend Snippet: Analyses of exosomes by FACS. (A) Exoxomes are CD63, LAMP2, CD81 and C3 positive, but CFH negative. Isotype control: Black; CFH: light green; CD63: blue; LAMP2: red; CD81: dark green; C3: magenta. (B) Added CFH bound to exosomes in a dose-dependent manner. (C) Added CFH did not alter the amount of C3 on exosomes. For B and C: CFH 0.5 mg/mL: black; CFH 0.05 mg/mL: blue; CFH 0.005 mg/mL: purple; CFH 0.0005 mg/mL: green; CFH 0 mg/mL: red.

    Article Snippet: 110 µl of 1 M glycine was added (i.e., 100 mM final), mixed gently and let stand on the bench at room temperature for 30 min. Beads were re-suspended in 0.5 ml PBS/0.5% BSA, 10 µl coated beads were incubated with 50 µl primary antibodies: mouse anti-CFH (1∶50, Serotec), mouse anti-CD63 (1∶50, Abcam), mouse ant-LAMP2 (1∶50, Abcam), mouse anti-CD81 (1∶50, Abcam), mouse anti-C3 (1∶50, Abcam).

    Techniques: FACS