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: Theranostics

Article Title: Exosome-mediated delivery of an anti-angiogenic peptide inhibits pathological retinal angiogenesis

doi: 10.7150/thno.54755

Figure Lengend Snippet: EXO KV11 suppresses VEGF-induced vascular leakage in vivo . (A) Schematic depiction of the pre-treatment procedure in a VEGF-induced vascular leakage model. EXO KV11 , KV11, EXO, or vehicle was retro-orbitally injected in adult wild type C57BL/6J mice. After 24 h, 100 ng VEGF was intravitreally injected to induce vascular leakage in the retina. The mice were sacrificed for analysis after another 24 h. (B) FITC-dextran was injected and the retinas were harvested at the endpoint. Representative images of flat-mounted retina show extravasated FITC-dextran and CD105 + vessels. (C) Quantification of dextran leakage in (B) (n = 7 - 10 mice per condition). (D) Representative confocal images of anti-CD105, anti-Claudin-5-stained retinal vessels in mice treated as in (A). (E) Representative images of F4/80 + macrophages (green) and CD105 + vessels in retinas treated as in (A). (F) Quantification of macrophage infiltration in (E) (n = 4 - 9 mice per condition). (G) Representative photographs of Evans blue leakage in SD rats ears in an auricular Miles assay with intradermal injection with VEGF, VEGF+KV11 mixture, or VEGF+EXO KV11 mixture. The data represent as mean ± SEM, *p < 0.05, **p < 0.01, ***p < 0.001, one-way ANOVA followed by Tukey's multiple comparisons test in (C), (F). Scale bars, 50 µm.

Article Snippet: For IF staining of red blood cells, blood vessels, macrophage and microglia, flat-mounted retinas were incubated in blocking solution with following primary antibodies at 4 °C overnight: rat anti-TER119 monoclonal (1:200, MAB1125, RD, US); rat anti-CD105 monoclonal (1:200, MAB1320, RD, US); rabbit anti-F4/80 monoclonal (1:200, 30325S, Cell Signaling Technology, US); mouse anti-Claudin-5 monoclonal (1:200, 35-2500, Invitrogen, US) and rabbit anti-ZO-1 polyclonal (1:300, 61-7300, Invitrogen, US).

Techniques: In Vivo, Injection, Staining

Journal: Theranostics

Article Title: Exosome-mediated delivery of an anti-angiogenic peptide inhibits pathological retinal angiogenesis

doi: 10.7150/thno.54755

Figure Lengend Snippet: EXO KV11 has therapeutic effect in post-treatment model. (A) Schematic depiction of the post-treatment procedure in VEGF-induced vascular leakage model. 100 ng VEGF was intravitreally injected to induce vascular leakage in the retina. After 24 h, EXO KV11 , KV11, EXO, or vehicle was retro-orbitally injected in adult wild type C57BL/6J mice. The mice were sacrificed for analysis after another 48 h. (B) FITC-dextran was injected and the retinas were harvested after treatment. Representative images of flat-mounted retina show extravasated FITC-dextran and CD105 + vessels. (C) Quantification of dextran leakage in (B) (n = 4 - 6 mice per condition). (D) Representative images of F4/80 + macrophages (green) and CD105 + vessels in retinas treated as in (A). (E) Quantification of macrophage infiltration in (D) (n = 4 - 6 mice per condition). The data represent as mean ± SEM. *p < 0.05, **p < 0.01, ***p < 0.001, one-way ANOVA followed by Tukey's multiple comparisons test in (C), (E). Scale bars, 50 µm.

Article Snippet: For IF staining of red blood cells, blood vessels, macrophage and microglia, flat-mounted retinas were incubated in blocking solution with following primary antibodies at 4 °C overnight: rat anti-TER119 monoclonal (1:200, MAB1125, RD, US); rat anti-CD105 monoclonal (1:200, MAB1320, RD, US); rabbit anti-F4/80 monoclonal (1:200, 30325S, Cell Signaling Technology, US); mouse anti-Claudin-5 monoclonal (1:200, 35-2500, Invitrogen, US) and rabbit anti-ZO-1 polyclonal (1:300, 61-7300, Invitrogen, US).

Techniques: Injection

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