Journal: Oncology reports

Article Title: A novel peptide targets CD105 for tumour imaging in vivo.

doi: 10.3892/or.2018.6643

Figure Lengend Snippet: Figure 2. (A) Flow cytometry analysis of the binding affinity of 13 peptides for MNNG/HOS cells. nABP296 and nABP297 peptides exhibited higher binding affinity for the MNNG/HOS cell line. (B) MFI analysis of the flow cytometry results, demonstrating that nABP296 and nABP297 had the highest MFI. (C) Flow cytometry analysis of nABP296 and nABP297 in MNNG/HOS and Cal27 cells. nABP297 exhibited no binding affinity difference between MNNG/HOS and Cal27 cell lines, while nABP296 had a higher binding affinity for MNNG/HOS cells and lower affinity for Cal27 cells. Thus, nABP296 could be used for MNNG/HOS labelling in vitro and in vivo. (D) Immunofluorescence analysis of nABP296 in MNNG/HOS and Cal27 cells. nABP296 was co‑located with the CD105 antibody in MNNG/HOS cells, whereas both CD105 and nABP296 exhibited no binding to Cal27. (E) ELISA of nABP296 binding affinity for CD105 protein, with CD106 protein used as control. The OD value in the CD105 protein group was higher than that in the CD106 protein group (*P<0.05 vs. control). (F) Cytotoxicity assay of nABP296 on MNNG/HOS cells at 24 and 48 h, exhibiting no cytotoxicity on the MNNG/HOS cell line (P>0.05). nABP, non‑antibody‑binding protein; MFI, mean fluorescence intensity; OD, optical density.

Article Snippet: Subsequent to collection in a 0.25% (w/v) trypsin solution with 0.025% (w/v) EDTA, MNNG/HOS and Cal27 cells were incubated with the CD105 antibody (cat. no. 130‐102‐819; 1:10; Miltenyi Biotec GmbH, Bergisch Gladbach, Germany) at 4 ̊C for 10 min.

Techniques: Flow Cytometry, Binding Assay, In Vitro, In Vivo, Immunofluorescence, Enzyme-linked Immunosorbent Assay, Control, Cytotoxicity Assay, Fluorescence

Journal: Oncology reports

Article Title: A novel peptide targets CD105 for tumour imaging in vivo.

doi: 10.3892/or.2018.6643

Figure Lengend Snippet: Figure 1. (A) Semiquantitative reverse transcription‑polymerase chain reaction analysis of CD105 levels in MNNG/HOS and Cal27 cell lines. CD105 mRNA expression was observed in MNNG/HOS cells, whereas there was no CD105 mRNA expression in Cal27 cells. (B) Immunofluorescence analysis of CD105 expression in MNNG/HOS and Cal27 cell lines. As shown by red fluorescence, CD105 was located on the surface of MNNG/HOS cells. There was no CD105 fluorescence detected in the Cal27 cell line. (C) Flow cytometry analysis of CD105 expression in MNNG/HOS and Cal27 cell lines, confirming that MNNG/HOS cells highly expressed CD105 protein, while this was not expressed in Cal27 cells.

Article Snippet: Subsequent to collection in a 0.25% (w/v) trypsin solution with 0.025% (w/v) EDTA, MNNG/HOS and Cal27 cells were incubated with the CD105 antibody (cat. no. 130‐102‐819; 1:10; Miltenyi Biotec GmbH, Bergisch Gladbach, Germany) at 4 ̊C for 10 min.

Techniques: Reverse Transcription Polymerase Chain Reaction, Expressing, Immunofluorescence, Fluorescence, Flow Cytometry

Journal: Oncology reports

Article Title: A novel peptide targets CD105 for tumour imaging in vivo.

doi: 10.3892/or.2018.6643

Figure Lengend Snippet: Figure 4. In vitro visualisation assay. (A) nABP296 exhibited colocalization with CD105 antibody in tumour sections derived from MNNG/HOS tumour‑bearing mice. (B) A tumour histological section derived from an osteosarcoma patient could be label by nABP296 peptide and CD105 antibody. (C) Tumour histological section derived from an oral carcinoma patient, in which there was no CD105 antibody fluorescence and low nABP296 fluorescence. nABP, non‑antibody‑binding protein.

Article Snippet: Subsequent to collection in a 0.25% (w/v) trypsin solution with 0.025% (w/v) EDTA, MNNG/HOS and Cal27 cells were incubated with the CD105 antibody (cat. no. 130‐102‐819; 1:10; Miltenyi Biotec GmbH, Bergisch Gladbach, Germany) at 4 ̊C for 10 min.

Techniques: In Vitro, Derivative Assay, Fluorescence

Journal: Stem Cells International

Article Title: Cancer Stem Cell Marker Endoglin (CD105) Induces Epithelial Mesenchymal Transition (EMT) but Not Metastasis in Clear Cell Renal Cell Carcinoma

doi: 10.1155/2019/9060152

Figure Lengend Snippet: CD105(+) ccRCC tumor cells are mesenchymal cells with enhanced motility and invasion capability. (a) Western blot and (b) qRT-PCR of EMT markers of CD105(+) cells and parental cells. (c) Migration assay and (d) modified 3D transwell assay of CD105(+) cells and parental cells ( ∗ p < 0.05, ∗∗ p < 0.01).

Article Snippet: For flow cytometry antibodies, anti-human CD105 (130-112-321) conjugated with PE antibody was bought from Miltenyi (CA, USA) and was used for regular verification of CD105 expression in CD105(+) cell within 50 passages.

Techniques: Western Blot, Quantitative RT-PCR, Migration, Modification, Transwell Assay

Journal: Stem Cells International

Article Title: Cancer Stem Cell Marker Endoglin (CD105) Induces Epithelial Mesenchymal Transition (EMT) but Not Metastasis in Clear Cell Renal Cell Carcinoma

doi: 10.1155/2019/9060152

Figure Lengend Snippet: CD105 knockdown induces loss of mesenchymal markers and inhibition of motility and invasion. (a) Western blot and (b) qRT-PCR of EMT markers of CD105(+) cells and shRNA-mediated CD105 knockdown CD105(+) cells. (c) Migration assay and (d) modified 3D transwell assay of CD105(+) cells and shRNA-mediated CD105 knockdown CD105(+) cells ( ∗ p < 0.05, ∗∗ p < 0.01).

Article Snippet: For flow cytometry antibodies, anti-human CD105 (130-112-321) conjugated with PE antibody was bought from Miltenyi (CA, USA) and was used for regular verification of CD105 expression in CD105(+) cell within 50 passages.

Techniques: Knockdown, Inhibition, Western Blot, Quantitative RT-PCR, shRNA, Migration, Modification, Transwell Assay

Journal: Stem Cells International

Article Title: Cancer Stem Cell Marker Endoglin (CD105) Induces Epithelial Mesenchymal Transition (EMT) but Not Metastasis in Clear Cell Renal Cell Carcinoma

doi: 10.1155/2019/9060152

Figure Lengend Snippet: MYC overexpression can reverse this process. (a) Western blot of MYC in CD105(+) cells, shRNA-mediated CD105 knockdown CD105(+) cells, and MYC overexpressed cells. (b) qRT-PCR of EMT markers of shRNA-mediated CD105 knockdown CD105(+) cells before and after MYC overexpression. (c) Migration assay of shRNA-mediated CD105 knockdown CD105(+) cells and MYC overexpressed cells. (d) Migration assay of shRNA-mediated CD105 knockdown CD105(+) cells and NANOG overexpressed cells. (e) qRT-PCR analysis of EMT markers of CD105(+) cells before and after the treatment of TGF- β type I receptor kinase inhibitor LY-364947 (50 nM) ( ∗∗ p < 0.01).

Article Snippet: For flow cytometry antibodies, anti-human CD105 (130-112-321) conjugated with PE antibody was bought from Miltenyi (CA, USA) and was used for regular verification of CD105 expression in CD105(+) cell within 50 passages.

Techniques: Over Expression, Western Blot, shRNA, Knockdown, Quantitative RT-PCR, Migration

Journal: Stem Cells International

Article Title: Cancer Stem Cell Marker Endoglin (CD105) Induces Epithelial Mesenchymal Transition (EMT) but Not Metastasis in Clear Cell Renal Cell Carcinoma

doi: 10.1155/2019/9060152

Figure Lengend Snippet: CD105 knockdown does not change the metastasis of ccRCC in the tail vein injection mouse model. (a) The lung weight analysis of mice received the CD105(+) cells and shRNA-mediated CD105 knockdown CD105(+) cells. Representative images of lung metastasis in both gross view (b) and H&E staining (c) established by tail vein injection of cancer cells.

Article Snippet: For flow cytometry antibodies, anti-human CD105 (130-112-321) conjugated with PE antibody was bought from Miltenyi (CA, USA) and was used for regular verification of CD105 expression in CD105(+) cell within 50 passages.

Techniques: Knockdown, Injection, shRNA, Staining

Journal: Cell

Article Title: Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches

doi: 10.1016/j.cell.2021.12.018

Figure Lengend Snippet:

Article Snippet: Anti-Mouse CD105 (REA1058) FITC , Miltenyi Biotec , 130-118-173; RRID: AB_2733613.

Techniques: Purification, Recombinant, Staining, cDNA Synthesis, Gene Expression, Software, Microscopy

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

Article Title: Targeting CD146 with a 64 Cu-labeled antibody enables in vivo immunoPET imaging of high-grade gliomas

doi: 10.1073/pnas.1502648112

Figure Lengend Snippet: CD146 clinical relevance in glioblastoma multiforme patients. Clinical data were obtained from TCGA. (A) The table summarizes the demographics of the analyzed patient cohort. A Kaplan–Meier plot showing a significant difference in (B) disease-free survival (DFS) and (C) overall survival between CD146(+) and CD146(−) GBM patients. P values were determined by the log-rank test.

Article Snippet: Real-time PCR was performed using TaqMan primers for CD146 (Mm00468256_m1 MCM) and GAPDH (Mm99999915_g1 Gapdh) (Applied Biosystems).

Techniques:

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

Article Title: Targeting CD146 with a 64 Cu-labeled antibody enables in vivo immunoPET imaging of high-grade gliomas

doi: 10.1073/pnas.1502648112

Figure Lengend Snippet: Modified methodology for the production of anti-human CD146 mAb. (A) Schematic representation of the mAb production workflow. A modified protocol was used for the immunization and isolation of activated B cells, which permitted a significant cut in the production time and cost. (B) Selection of the most immunoreactive mAb clones. The immunoreactivity of the antibodies was determined by immunofluorescence staining of human melanoma A375 cells, known to overexpress the CD146 antigen; YY146 (clone no. 5) displayed the strongest affinity for the CD146 antigen. (Scale bars: 50 µm.)

Article Snippet: Real-time PCR was performed using TaqMan primers for CD146 (Mm00468256_m1 MCM) and GAPDH (Mm99999915_g1 Gapdh) (Applied Biosystems).

Techniques: Modification, Isolation, Selection, Clone Assay, Immunofluorescence, Staining

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

Article Title: Targeting CD146 with a 64 Cu-labeled antibody enables in vivo immunoPET imaging of high-grade gliomas

doi: 10.1073/pnas.1502648112

Figure Lengend Snippet: In vitro screenings for antibody clone selection. (A) Illustration of the ELISAs for each of the purified mAb clones. (B) Immunoblotting of CD146 showing the antigen recognition by the five antibody clones and a commercially available anti-CD146 mAb. (C) CD146 immunofluorescence staining of A375 cells using three of the generated clones (II, III, and YY146); YY146 showed a markedly higher staining of the cell membranes. E9653 is a commercially available anti-CD146 mAb.

Article Snippet: Real-time PCR was performed using TaqMan primers for CD146 (Mm00468256_m1 MCM) and GAPDH (Mm99999915_g1 Gapdh) (Applied Biosystems).

Techniques: In Vitro, Selection, Purification, Clone Assay, Western Blot, Immunofluorescence, Staining, Generated

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

Article Title: Targeting CD146 with a 64 Cu-labeled antibody enables in vivo immunoPET imaging of high-grade gliomas

doi: 10.1073/pnas.1502648112

Figure Lengend Snippet: In vitro characterization of CD146 expression in human cancer cells. (A) Flow cytometry of U87MG and U251 human glioblastoma cells. U87MG cells showed markedly higher fluorescence signal, indicative of stronger YY146 binding than U251 cells. (B) Immunofluorescence staining of CD146 in U87MG and U251 cells and tissues. Consistently with A, U87MG cells and tissue show stronger staining than U251 specimens.

Article Snippet: Real-time PCR was performed using TaqMan primers for CD146 (Mm00468256_m1 MCM) and GAPDH (Mm99999915_g1 Gapdh) (Applied Biosystems).

Techniques: In Vitro, Expressing, Flow Cytometry, Fluorescence, Binding Assay, Immunofluorescence, Staining

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

Article Title: Targeting CD146 with a 64 Cu-labeled antibody enables in vivo immunoPET imaging of high-grade gliomas

doi: 10.1073/pnas.1502648112

Figure Lengend Snippet: Conjugation of NOTA to YY146 does not affect its immunoreactivity. (A) FACS analysis of U87MG (CD146+) and U251 (CD146−) showing fluorescence signal with both YY146 and NOTA-YY146. (B) SDS/PAGE of both YY146 and NOTA-YY146 revealed no significant difference in molecular mass. (C) Immunofluorescence staining of U87MG cells displayed very similar staining patterns. (D) Assessment of NOTA-YY146 serum stability, in vitro. Incubation of NOTA-YY146 with mouse serum for up to 48 h did not compromise the immunoreactivity of the antibody.

Article Snippet: Real-time PCR was performed using TaqMan primers for CD146 (Mm00468256_m1 MCM) and GAPDH (Mm99999915_g1 Gapdh) (Applied Biosystems).

Techniques: Conjugation Assay, Fluorescence, SDS Page, Immunofluorescence, Staining, In Vitro, Incubation

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

Article Title: Targeting CD146 with a 64 Cu-labeled antibody enables in vivo immunoPET imaging of high-grade gliomas

doi: 10.1073/pnas.1502648112

Figure Lengend Snippet: Noninvasive PET imaging of the in vivo CD146 expression and ex vivo biodistribution of 64Cu-NOTA-YY146 in mice bearing s.c. glioblastoma xenografts. (A) Sequential PET images of coronal planes containing s.c. U87MG or U251 tumors. (Top) Elevated uptake of the tracer in the U87MG tumor xenografts. Lower tumor uptake was observed in low CD146 expressing U251 tumors (Middle) or when U87MG tumor-bearing mice were preinjected with a blocking dose of the unlabeled antibody (Bottom). H, heart; L, liver; S, spleen, T, tumor. (B) Region of interest (ROI) analysis of the PET images at 4 h, 24 h, and 48 h p.i. showing 64Cu-NOTA-YY146 uptake values in blood pool, liver, muscle, and tumor; values are presented as %ID/g ± SD (n = 3 or 4). Tracer accumulation was significantly higher in U87MG xenografts compared with U251 or blocked U87MG groups. (C) Ex vivo biodistribution data 48 h after inoculation of the tracer. Mice were euthanized immediately after the last PET scan, and the tissues were collected, weighed, and counted; the results were presented as %ID/g ± SD (n = 3 or 4).

Article Snippet: Real-time PCR was performed using TaqMan primers for CD146 (Mm00468256_m1 MCM) and GAPDH (Mm99999915_g1 Gapdh) (Applied Biosystems).

Techniques: Imaging, In Vivo, Expressing, Ex Vivo, Blocking Assay

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

Article Title: Targeting CD146 with a 64 Cu-labeled antibody enables in vivo immunoPET imaging of high-grade gliomas

doi: 10.1073/pnas.1502648112

Figure Lengend Snippet: CD146/CD31 immunofluorescence staining of U87MG, U251, and muscle tissue. U87MG tissue showed a pronounced CD146 staining whereas only background levels were seen in U251 or muscle tissues.

Article Snippet: Real-time PCR was performed using TaqMan primers for CD146 (Mm00468256_m1 MCM) and GAPDH (Mm99999915_g1 Gapdh) (Applied Biosystems).

Techniques: Immunofluorescence, Staining

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

Article Title: Targeting CD146 with a 64 Cu-labeled antibody enables in vivo immunoPET imaging of high-grade gliomas

doi: 10.1073/pnas.1502648112

Figure Lengend Snippet: Histological analysis of glioblastoma tumors obtained from mice bearing MRI-confirmed orthotopic xenografts. T2-weighted MRI shows a distinctive signal enhancement corresponding to the tumor mass. Whole brain coronal tissue sections containing the tumors were prepared for H&E and immunofluorescence (IFS) staining. H&E staining (Left) shows the distinctive morphological traits of U87MG and U251 tumors compared with the surrounding normal brain. CD146/CD31 IFS (Right) of brain sections revealed a markedly higher CD146 expression (green color) in U87MG tissue whereas U251 and the adjacent normal tissue displayed only low background staining. (Scale bars: 50 µm.)

Article Snippet: Real-time PCR was performed using TaqMan primers for CD146 (Mm00468256_m1 MCM) and GAPDH (Mm99999915_g1 Gapdh) (Applied Biosystems).

Techniques: Immunofluorescence, Staining, Expressing

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

Article Title: Targeting CD146 with a 64 Cu-labeled antibody enables in vivo immunoPET imaging of high-grade gliomas

doi: 10.1073/pnas.1502648112

Figure Lengend Snippet: PET/CT imaging and ex vivo biodistribution studies of 64Cu-NOTA-YY146 in mice bearing orthotopic glioblastoma xenografts. (A) Representative sequential PET/CT images of coronal and sagittal planes containing the U87MG or U251 brain tumors. Images revealed a marked accumulation of 64Cu-NOTA-YY146 within U87MG tumor at 4 h, 24 h, and 48 h after injection. (B) Comparison of PET ROI-derived quantification of 64Cu-NOTA-YY146 uptake in U87MG (CD146+) vs. U251 (CD146−) tumors. Significantly higher accretion of the tracer in U87MG tumors was observed at 4-h, 24-h, and 48-h time points; reported as %ID/g ± SD (n = 5). (C) Ex vivo biodistribution of 64Cu-NOTA-YY146 in tumor-bearing mice, 48 h p.i. Uptake in several of the major organs/tissues is presented as %ID/g ± SD (n = 4).

Article Snippet: Real-time PCR was performed using TaqMan primers for CD146 (Mm00468256_m1 MCM) and GAPDH (Mm99999915_g1 Gapdh) (Applied Biosystems).

Techniques: Positron Emission Tomography-Computed Tomography, Imaging, Ex Vivo, Injection, Derivative Assay

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

Article Title: Targeting CD146 with a 64 Cu-labeled antibody enables in vivo immunoPET imaging of high-grade gliomas

doi: 10.1073/pnas.1502648112

Figure Lengend Snippet: Flow cytometry analysis of the expression of CD146, CD44, and CD133 in U87MG cells. (A) Control dot plot. (B) Representative flow cytometry dot plot depicting CD146-positive and CD133-negative expression in U87MG cells. (C) Dot plot showing the coexpression of both CD44 and CD146.

Article Snippet: Real-time PCR was performed using TaqMan primers for CD146 (Mm00468256_m1 MCM) and GAPDH (Mm99999915_g1 Gapdh) (Applied Biosystems).

Techniques: Flow Cytometry, Expressing

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

Article Title: Targeting CD146 with a 64 Cu-labeled antibody enables in vivo immunoPET imaging of high-grade gliomas

doi: 10.1073/pnas.1502648112

Figure Lengend Snippet: YY146 preferentially targets stem cell-like tumor cells and reverts their mesenchymal phenotypes. (A) Western blot of nonsorted, CD146 low (+) and CD146 high (++) U87MG cell populations. Using FACS, U87MG cells were separated into two population, U87MG(+) and U87MG(++), and the correlation between CD146 levels and the expression of several markers associated with stem cell and mesenchymal phenotypes was determined. The U87MG(++) population displayed marked expression signatures corresponding to stem cell and mesenchymal phenotype. (B) RT-PCR of U87MG cells treated with 10 µg/mL YY146 for 0, 6, 12, and 24 h. A decline on the mRNA levels of CD146 and RAC1 was observed whereas E-cadherin was up-regulated upon YY146 treatment. (C) Western blots of the YY146-treated cells. Variation on the mRNA levels of CD146, E-cadherin, and RAC1 translated into similar variations on the corresponding protein expression; additionally, a reduction in the expression of Nanog was observed. (D) A transwell invasion assay revealed a significant decline in the invasive and migratory behavior of U87MG cells after treatment with YY146, but not in the isotype-matched antibody or vehicle controls. (E) YY146 toxicity in U87MG cells. High cell survival was appreciated even after prolonged incubation (48 h) of cells with 1 mg/mL YY146.

Article Snippet: Real-time PCR was performed using TaqMan primers for CD146 (Mm00468256_m1 MCM) and GAPDH (Mm99999915_g1 Gapdh) (Applied Biosystems).

Techniques: Western Blot, Expressing, Reverse Transcription Polymerase Chain Reaction, Transwell Invasion Assay, Incubation

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

Article Title: Targeting CD146 with a 64 Cu-labeled antibody enables in vivo immunoPET imaging of high-grade gliomas

doi: 10.1073/pnas.1502648112

Figure Lengend Snippet: CD146 expression is associated with human brain cancer histological grade. (A) Summary of glioma patient demographics and their corresponding CD146 expression levels. (B) IHC staining of CD146 in different WHO grade gliomas. CD146 staining of grades III and IV malignancies is markedly higher (dark brown color) compared with grade I and II tumors.

Article Snippet: Real-time PCR was performed using TaqMan primers for CD146 (Mm00468256_m1 MCM) and GAPDH (Mm99999915_g1 Gapdh) (Applied Biosystems).

Techniques: Expressing, Immunohistochemistry, Staining

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

Article Title: Targeting CD146 with a 64 Cu-labeled antibody enables in vivo immunoPET imaging of high-grade gliomas

doi: 10.1073/pnas.1502648112

Figure Lengend Snippet: CD146 is overexpressed in a wide range of malignancies. (A) In vitro FACS analysis of CD146 expression in 10 different cancer cell lines using YY146 as the primary antibody, many of which express a high level of CD146. (B) IHC analysis showing CD146 expression in several human primary tumors (using YY146 as the primary antibody). A markedly stronger CD146 staining is observed in high-grade malignancies. (C) Semiquantitative analysis of the histological data revealed a clear tendency of tumors of higher grade to express a higher level of CD146 than lower grade tumors.

Article Snippet: Real-time PCR was performed using TaqMan primers for CD146 (Mm00468256_m1 MCM) and GAPDH (Mm99999915_g1 Gapdh) (Applied Biosystems).

Techniques: In Vitro, Expressing, Staining

Journal: Molecular Neurodegeneration

Article Title: Endothelium-specific endoglin triggers astrocyte reactivity via extracellular vesicles in a mouse model of Alzheimer’s disease

doi: 10.1186/s13024-025-00875-4

Figure Lengend Snippet: CEEVs specific endoglin (ENG) is a potential communicator between BMECs and astrocytes in AD. a Schematic and timeline of the transplantation treatment by PKH26-labeled cerebrovascular endothelial extracellular vesicles (CEEVs). b , c The bio-distribution of CEEVs-PKH26 in vivo at 24 h ( b ) and in body tissues 48 h after injection ( n = 3 mice per group) ( c ). d-g Representative images ( d ), the quantification of PKH26 fluorescence intensity in hippocampal dentate gyrus ( n = 3 mice per group) ( e ), the percentage of PKH26 distributed in GFAP staining ( n = 3 mice per group) ( f ), and the quantification of GFAP per astrocyte cell with or without CEEVs from 3 mice per group ( g ). h Differential-gene expression analysis of HCMEC/D3 stimulated by Ang II. Dotted lines indicate HCMEC-DEGs cut-offs for |log2(fold change)|>1.5 and -log10 ( P -value) of 1.3, corresponding to P -value < 0.05, 3 samples per group. i The number of specific genes of HCMEC-DEGs in endothelial cells compared with other cell types in brain (microglia, neuron, astrocyte and oligodendrocyte). j Venn diagrams showing the identification of ENG by intersecting the DEGs of injured BMECs from ( h ) with the differential proteome in the CSF of AD patients and the proteome of endothelial cell EVs from the datasets. k-m A cohort study of AD patients ( n = 18) and non-demented control ( n = 15). ENG protein levels were up-regulated in the serum of AD patients measured by ELISA ( k ). Scatter plot of Montreal Cognitive Assessment (MoCA) versus ENG levels was shown and the data were analyzed with a linear regression method ( l ) and ROC curve analysis of ENG ( m ). Data are shown as means ± SEM, and the P -value was reported on the graph highlighted comparison by unpaired two-sided t -test ( c , e , f , k ) and one-way ANOVA with post-hoc Tukey adjustment ( g ). ENG: Endoglin, ECs: endothelial cells, DEGs: differential genes, NC: Non-demented control

Article Snippet: Mice primary astrocytes were treated with recombinant human ENG protein (SinoBiological, #10149-H02H, China) at 50 ng/mL or SB431542 (MedChemExpress, #HY-10431, USA) at 10 μM for 24 h for qRT-PCR or Western blotting.

Techniques: Transplantation Assay, Labeling, In Vivo, Injection, Fluorescence, Staining, Gene Expression, Control, Enzyme-linked Immunosorbent Assay, Comparison

Journal: Molecular Neurodegeneration

Article Title: Endothelium-specific endoglin triggers astrocyte reactivity via extracellular vesicles in a mouse model of Alzheimer’s disease

doi: 10.1186/s13024-025-00875-4

Figure Lengend Snippet: Injured BMECs deliver ENG protein to astrocytes through CEEVs. a , b Representative images ( a ) and the quantification ( b ) of ENG on per astrocyte in WT and APP/PS1 mice ( n = 5 mice per group). c ENG protein levels in HCMEC/D3 conditioned-medium incubating with or without astrocytes ( n = 6 biologically independent experiments). d , e Immunostaining ( d ) and the quantification ( e ) of ENG expression in HCMEC/D3 treated with Ang II ( n = 6 biologically independent experiments). f ENG protein levels in HCMEC/D3 medium treated by Ang II ( n = 6 biologically independent experiments). g Immunoblotting of distribution of ENG in HCMEC/D3 culture medium. h Protein content of ENG in the CEEVs from Ang II-treated HCMEC/D3 vs. normal control ( n = 3 biologically independent experiments). i , j Representative images ( i ) and the quantification ( j ) of ENG spots on per astrocyte cell in AST & HC Co-culture system treated by GW4869 to HCMEC/D3 or Amiloride to astrocytes from 3 biologically independent experiments. Data are shown as means ± SEM, and the P -value was reported on the graph highlighted comparison by unpaired two-sided t -test ( b , e ), two-way ANOVA with post-hoc Tukey adjustment ( c ), one-way ANOVA with post-hoc Tukey adjustment ( f ) and Kruskal-Wallis with Dunn’s multiple comparisons test ( j ). AST: astrocytes, HC: HCMEC/D3

Article Snippet: Mice primary astrocytes were treated with recombinant human ENG protein (SinoBiological, #10149-H02H, China) at 50 ng/mL or SB431542 (MedChemExpress, #HY-10431, USA) at 10 μM for 24 h for qRT-PCR or Western blotting.

Techniques: Immunostaining, Expressing, Western Blot, Control, Co-Culture Assay, Comparison

Journal: Molecular Neurodegeneration

Article Title: Endothelium-specific endoglin triggers astrocyte reactivity via extracellular vesicles in a mouse model of Alzheimer’s disease

doi: 10.1186/s13024-025-00875-4

Figure Lengend Snippet: CEEVs switch astrocyte reactivity through ENG. a Immunoblotting and the quantification of GFAP in astrocytes treated by ENG recombinant protein ( n = 3 biologically independent experiments). b Immunoblotting of GFAP in astrocytes treated by ENG-overexpressed CEEVs ( n = 3/8 biologically independent experiments). c , d Knockdown of ENG in HCMEC/D3 and immunoblotting of ENG and GFAP in astrocytes treated by ENG-knockdown CEEVs ( n = 3 biologically independent experiments). e-g Representative images ( e ) and the quantification of GFAP in per astrocyte ( f ) and PKH26 fluorescence intensity ( g ) in the hippocampus of CEEVs-treated C57BL/6J mice administrated with or without Carotuximab from 5 mice per group. Data are shown as means ± SEM, and the P -value was reported on the graph highlighted comparison by unpaired two-sided t -test

Article Snippet: Mice primary astrocytes were treated with recombinant human ENG protein (SinoBiological, #10149-H02H, China) at 50 ng/mL or SB431542 (MedChemExpress, #HY-10431, USA) at 10 μM for 24 h for qRT-PCR or Western blotting.

Techniques: Western Blot, Recombinant, Knockdown, Fluorescence, Comparison

Journal: Molecular Neurodegeneration

Article Title: Endothelium-specific endoglin triggers astrocyte reactivity via extracellular vesicles in a mouse model of Alzheimer’s disease

doi: 10.1186/s13024-025-00875-4

Figure Lengend Snippet: ENG regulates reactive astrocytes via TGFBRI/Smad3 pathway. a Immunoprecipitation and immunoblot analysis of ENG and TGFBRI in astrocytes of AST & HC Co-culture system. b Representative images and co-localization analysis of ENG and TGFBRI on astrocytes of AST & HC co-culture system. c Immunoblotting analysis and quantification of Smad3 and phospho-Smad3 (Ser423/425) in primary astrocytes treated with ENG recombinant protein ( n = 3 biologically independent experiments). d The levels of IL-6, IL-3 and VEGF in the medium of ENG-treated astrocytes ( n = 6 biologically independent experiments). e mRNA levels of GFAP and VIM in astrocytes treated by ENG ( n = 6 biologically independent experiments). f Relative mRNA levels of VIM in astrocytes treated by ENG or SB431542 ( n = 5 biologically independent experiments). Data are shown as means ± SEM, and the P -value was reported on the graph highlighted comparison by unpaired two-sided t -test ( c-e ) or one-way ANOVA with post-hoc Tukey adjustment ( f )

Article Snippet: Mice primary astrocytes were treated with recombinant human ENG protein (SinoBiological, #10149-H02H, China) at 50 ng/mL or SB431542 (MedChemExpress, #HY-10431, USA) at 10 μM for 24 h for qRT-PCR or Western blotting.

Techniques: Immunoprecipitation, Western Blot, Co-Culture Assay, Recombinant, Comparison

Journal: Molecular Neurodegeneration

Article Title: Endothelium-specific endoglin triggers astrocyte reactivity via extracellular vesicles in a mouse model of Alzheimer’s disease

doi: 10.1186/s13024-025-00875-4

Figure Lengend Snippet: ENG deficiency inhibits astrocyte reactivity and neuroinflammation in APP/PS1. a Schematic of APP/PS1 experimental procedure of AAV-shENG interference. b Western blotting to identify ENG expression in the hippocampus of AAV-shENG injected APP/PS1 mice ( n = 3 mice per group). c-e Representative images ( c ) and quantification of Smad3 ( d ) and GFAP ( e ) in the hippocampal dentate gyrus of APP/PS1-shCon and APP/PS1-shENG mice ( n = 3 mice per group). f mRNA levels of Serpina3N , VIM and Ctsb in the hippocampus of APP/PS1-shCon and APP/PS1-shENG mice ( n = 6 mice per group). g , h Representative images and quantification of VIM ( g ) and Aβ plaque ( h ) in the hippocampal dentate gyrus of APP/PS1-shCon and APP/PS1-shENG mice ( n = 3 mice per group). i Sholl analysis of astrocytes from the VIM-stained images. Interval of concentric circles is 3 μm. j Average number of intersections in Sholl analysis with respect to the distance from the center of each VIM-stained cell. k , l The sum number of intersects ( k ) and the ending radius ( l ) by Sholl analysis of the stained VIM signal of each VIM-stained cell from 3 mice per group. m Relative mRNA levels of IL-6 , IL-3 and VEGF in the hippocampus of APP/PS1-shCon and APP/PS1-shENG mice ( n = 6 mice per group). Data are shown as means ± SEM, and the P -value was reported on the graph highlighted comparison by unpaired two-tailed t -test

Article Snippet: Mice primary astrocytes were treated with recombinant human ENG protein (SinoBiological, #10149-H02H, China) at 50 ng/mL or SB431542 (MedChemExpress, #HY-10431, USA) at 10 μM for 24 h for qRT-PCR or Western blotting.

Techniques: Western Blot, Expressing, Injection, Staining, Comparison, Two Tailed Test