Journal:

Article Title: Comprehensive Detection and Serotyping of Human Adenoviruses by PCR and Sequencing

doi: 10.1128/JCM.42.9.3963-3969.2004

Figure Lengend Snippet: Adenovirus strains used in this study a

Article Snippet: 20 , ATCC VR-1097 , 626† , {"type":"entrez-nucleotide","attrs":{"text":"AY288110","term_id":"33637722","term_text":"AY288110"}} AY288110 †.

Techniques: Amplification

Journal: International Journal of Biological Sciences

Article Title: N-acetylcysteine and raloxifene boost photodynamic therapy against cutaneous squamous cell carcinoma by decreasing TGFβ1 secreted by cancer-associated fibroblasts

doi: 10.7150/ijbs.106642

Figure Lengend Snippet: Effect of CAF endoglin expression levels on TGFβ1 secretion and on the induction of resistance to PDT in A431 cells. (A) Secretion of TGFβ1 produced by T205A after treatment with increasing concentrations of recombinant endoglin. ELISA assay was performed to quantify the levels of secreted TGFβ1 to the culture medium collected 24 h after treatments. (B) Expression of endoglin analyzed by Western blot in T205A CAFs after transfection with the negative control (NC) and a siRNA targeting endoglin. Western blot images are representative of n = 3 independent experiments. Error bars denote ± S.E.M. (n = 4, one-way ANOVA: *p<0.05, **p<0.01 and ***p<0.001). (C) Secretion of TGFβ1 produced by T205A CAFs after transfection with NC or ENGsiRNA. ELISA assay was performed to quantify the levels of secreted TGFβ1 to the culture medium collected 24 h after treatment with NAC (D) Effect of the pre-treatment with NAC of transfected T205A in the CAF-derived CM on the response of bidimensional A431 cell cultures to PDT. Upper row: phase contrast images illustrating the morphological changes in A431 cells after treatments. Scale bar: 50 µm. Lower row: cell viability rates of A431 cells treated with MAL-PDT (0.5 mM of MAL and red light dose of 9.1 J·cm -2 ) in the presence of T205A CM after transfection. Error bars denote ± S.E.M. (n = 3, one-way ANOVA, statistical comparisons to Fb condition: *<0.05, **p<0.01 and ***p<0.001; multiple comparisons: #p<0.05, ##p<0.01 and ###p<0.001).

Article Snippet: Lyophilized recombinant human endoglin (R&D Systems, Minneapolis, MN, USA) was reconstituted in PBS containing 0.1% BSA to prepare a 250 μg/ml stock solution.

Techniques: Expressing, Produced, Recombinant, Enzyme-linked Immunosorbent Assay, Western Blot, Transfection, Negative Control, Derivative Assay

Journal: Cells

Article Title: Microglial Phenotyping in Neurodegenerative Disease Brains: Identification of Reactive Microglia with an Antibody to Variant of CD105/Endoglin

doi: 10.3390/cells8070766

Figure Lengend Snippet: Details of antibodies used in study.

Article Snippet: CD105 antibodies from R&D Systems (Minneapolis, MN, USA) (MAB1097) or Abcam (Cambridge, MA, USA.) (AB221675) were mixed for 18 hours with recombinant CD105 protein (1097-EN, R&D Systems), which consists of amino acids 26–586 of human CD105 expressed in mouse myeloma cells.

Techniques:

Journal: Cells

Article Title: Microglial Phenotyping in Neurodegenerative Disease Brains: Identification of Reactive Microglia with an Antibody to Variant of CD105/Endoglin

doi: 10.3390/cells8070766

Figure Lengend Snippet: Patterns of Staining of CD105/MAB1097 in human brain sections. ( a ) Staining of parallel sections of MTG from a low-plaque ND case for CD105/MAB1097 (panel A) (CD105/RnD) and IBA-1 (panel B). Staining of parallel sections of MTG from an AD case for CD105/RnD (panel C) or IBA-1 (panel D). All images have the same magnification. Scale bars represent 50 μm. ( b ) Low magnification images showing distribution of CD105-positive cells in low-plaque ND (panel A) and AD cases (panel B). Representative positive cells indicated by purple arrows. Both images at same magnification. Scale bars represent 100 μm. ( c ) Representative image of CD105 positive cells in low-plaque ND (LP) (panel A) (purple arrow) Morphology of CD105-positive microglia compared to IBA-1-positive surrounding cells (brown arrow) in LP case (panel B). Images of CD105-positive microglia with activated morphology in HP (panel C) and AD cases (panel D) (purple arrows). (panel E) Vascular staining in AD case (purple arrow) using CD105/Thermo Fisher antibody. All images at same magnification. Scale bars represent 50 μm.

Article Snippet: CD105 antibodies from R&D Systems (Minneapolis, MN, USA) (MAB1097) or Abcam (Cambridge, MA, USA.) (AB221675) were mixed for 18 hours with recombinant CD105 protein (1097-EN, R&D Systems), which consists of amino acids 26–586 of human CD105 expressed in mouse myeloma cells.

Techniques: Staining

Journal: Cells

Article Title: Microglial Phenotyping in Neurodegenerative Disease Brains: Identification of Reactive Microglia with an Antibody to Variant of CD105/Endoglin

doi: 10.3390/cells8070766

Figure Lengend Snippet: Biochemical Characterization of CD105 antibody MAB1097-detected polypeptides. ( a ) CD105 antibody MAB1097 (CD105/RnD)-identified polypeptides in protein samples of control unstimulated (CON), and polyIC (pIC) and Aβ-stimulated human brain-derived microglia (MG), and brain-derived endothelial cells (EC). ( b ) CD105 antibody MAB1097 (CD105/RnD)-identified polypeptides in protein samples of brain from middle temporal gyrus of high-plaque (HP), Alzheimer’s disease (AD) and low-plaque non-demented (ND) cases. ( c ) Antibody absorption studies. (Left hand panels): Images of Western blots containing brain MTG samples (LP, AD) and cell samples from THP-1-derived macrophages (MAC) reacted with CD105 antibody MAB1097 (RnD) preabsorbed with CD105 peptide (+Pep) or without peptide (-Pep). (Right hand panels): Images of Western blots of cell samples from THP-1-derived macrophages (MAC) or brain endothelial cell line hCMEC/D3 (EC) reacted with CD105 antibody AbCAM (AB221675) preabsorbed with CD105 peptide (+Pep) or without peptide (−Pep). ( d ) CD105 immunoreactive polypeptides after deglycosylation. Images of Western blots with CD105 antibodies MAB1097 (RnD), AB221675 (AbCAM) and PA5-32303 (Thermo) for two separate AD brain-derived protein extracts (AD1 and AD2) treated with PNGase F enzyme (+) or without enzyme (−). ( e ) Immunoprecipitation with CD105 antibodies. Images of Western blots with samples of brain (AD1 and AD2) or THP-1-derived macrophages (MAC) samples immunoprecipated with CD105 antibodies AbCAM or Thermo Fisher, and detected with CD105/RnD. MAC-T: sample of total protein of THP-1 macrophages.

Article Snippet: CD105 antibodies from R&D Systems (Minneapolis, MN, USA) (MAB1097) or Abcam (Cambridge, MA, USA.) (AB221675) were mixed for 18 hours with recombinant CD105 protein (1097-EN, R&D Systems), which consists of amino acids 26–586 of human CD105 expressed in mouse myeloma cells.

Techniques: Control, Derivative Assay, Western Blot, Immunoprecipitation

Journal: Cells

Article Title: Microglial Phenotyping in Neurodegenerative Disease Brains: Identification of Reactive Microglia with an Antibody to Variant of CD105/Endoglin

doi: 10.3390/cells8070766

Figure Lengend Snippet: Absorption controls for CD105 staining of microglia and antibody characterization. ( a ) Antibody absorption. (panels A–D). Staining of sections from AD cases (AD1) (A and B) or AD2 with CD105 antibody preabsorbed with immunizing peptide (+Pep) compared to staining of matched sections with CD105 antibody non-absorbed (−Pep). All images are at same magnification: scale bars represent 50 μm. ( b ) Deglycosylation. Tissue section (AD case) was treated with PNGase F in solution (+PNGase F) (panel A) or underwent control treatment without enzyme (−PNGase F) (panel B), and then reacted with CD105/RnD antibody. Images are at same magnification: scale bars represent 50 μm. ( c ) Antigen retrieval. Tissue sections from an AD case underwent antigen retrieval procedure prior to immunocytochemistry with antibodies CD105/RnD (panel A) or CD105/AbCAM (panel B). The position of an unstained large vessel is outlined (panel A). Vessel staining indicated with purple arrows (panel B). Scale bars represent 50 μm.

Article Snippet: CD105 antibodies from R&D Systems (Minneapolis, MN, USA) (MAB1097) or Abcam (Cambridge, MA, USA.) (AB221675) were mixed for 18 hours with recombinant CD105 protein (1097-EN, R&D Systems), which consists of amino acids 26–586 of human CD105 expressed in mouse myeloma cells.

Techniques: Staining, Control, Immunocytochemistry

Journal: Cells

Article Title: Microglial Phenotyping in Neurodegenerative Disease Brains: Identification of Reactive Microglia with an Antibody to Variant of CD105/Endoglin

doi: 10.3390/cells8070766

Figure Lengend Snippet: Images demonstrating colocalization of CD105/MAB1097 with microglial markers ( a ) panels A, B Low magnification images of high-plaque (HP) case (panel A) and AD case (panel B) showing staining with CD105/MAB1097 (purple arrows) and IBA-1 (brown arrows) in MTG sections. Both images at same magnification: scale bars represent 100 μm. ( b ) (panel A) CD105 microglia (purple arrows) with ramified morphology stained with antibody to P2RY12 (brown arrow) in low-plaque (LP) case. (panel B) CD105 microglia (purple arrows) and HLA-DR (brown arrow) in AD case. (panel C) CD105 microglia (purple arrow) and CD45 (brown arrow) in AD case. All sections were from MTG. Images at same magnification: scale bars represent 50 μm. ( c ) (panel A–E). Patterns of staining of CD105 (purple) and IBA-1 (brown) in MTG sections of cases with progressively increasing pathology from LP to AD. (panel F). CD105-positive microglia in hippocampus (HPC) of AD case. Images at same magnification: scale bars represent 50 μm.

Article Snippet: CD105 antibodies from R&D Systems (Minneapolis, MN, USA) (MAB1097) or Abcam (Cambridge, MA, USA.) (AB221675) were mixed for 18 hours with recombinant CD105 protein (1097-EN, R&D Systems), which consists of amino acids 26–586 of human CD105 expressed in mouse myeloma cells.

Techniques: Staining

Journal: Cells

Article Title: Microglial Phenotyping in Neurodegenerative Disease Brains: Identification of Reactive Microglia with an Antibody to Variant of CD105/Endoglin

doi: 10.3390/cells8070766

Figure Lengend Snippet: Different patterns of colocalization of CD105/MAB1097 and IBA-1 in microglia in AD cases by confocal microscopy. ( a ) IBA-1-positive (green) microglia; CD105-positive immunoreactive cells (red); Merged image showing colocalized CD105 and IBA-1 (yellow). Blue identifies nuclei revealed by staining with DAPI. Scale bars represent 50 μm. ( b ) Higher magnification image of a single cell with activated morphology. IBA-1-positive (green) microglia, strongly staining CD105-positive microglia with activated morphology (red). Merged image showing colocalized signals (yellow) within cell. Scale bars represent 25 μm. ( c ) Images to show different amounts of CD105 expression in microglia in an AD case. CD105 immunoreactivity (green), IBA-1 immunoreactivity (red), and merged image (yellow). Scale bars represent 50 μm.

Article Snippet: CD105 antibodies from R&D Systems (Minneapolis, MN, USA) (MAB1097) or Abcam (Cambridge, MA, USA.) (AB221675) were mixed for 18 hours with recombinant CD105 protein (1097-EN, R&D Systems), which consists of amino acids 26–586 of human CD105 expressed in mouse myeloma cells.

Techniques: Confocal Microscopy, Staining, Expressing

Journal: Cells

Article Title: Microglial Phenotyping in Neurodegenerative Disease Brains: Identification of Reactive Microglia with an Antibody to Variant of CD105/Endoglin

doi: 10.3390/cells8070766

Figure Lengend Snippet: Features of CD105/MAB1097-positive microglia in substantia nigra. ( a ) Control (non-PD case). (panel A) Association of CD105-positive microglia (purple arrows) and IBA-1 positive microglia (brown arrows) with substantia nigra dopaminergic neuromelanin-containing neurons. (panel B) Higher magnification showing interaction of CD105-positive microglia (purple) with degenerating neuromelanin containing dopaminergic neurons. The area imaged in B) is indicated by boxed area in A). Scale bars represent 50 μm. ( b ) ILBD case). (panel A) Association of CD105-positive microglia (purple arrows) and IBA-1-positive microglia (brown arrows) with substantia nigra dopaminergic neuromelanin-containing neurons showing greater numbers of CD105/RnD positive microglia. (panel B) Higher magnification showing greater interaction of CD105-positive microglia (purple arrows) with degenerating neuromelanin-containing dopaminergic neurons and free neuromelanin. Examples of some neuromelanin structures indicated by brown arrowheads. The area of image in B) is indicated by boxed area on A). Scale bars represent 50 υm. ( c ) PD cases. Sparse numbers of CD105-positive microglia (purple arrows) associated with surviving dopaminergic neurons in two separate PD cases panel A and B. The inset in panel B represents the boxed area at higher magnification. Scale bars represent 50 μm).

Article Snippet: CD105 antibodies from R&D Systems (Minneapolis, MN, USA) (MAB1097) or Abcam (Cambridge, MA, USA.) (AB221675) were mixed for 18 hours with recombinant CD105 protein (1097-EN, R&D Systems), which consists of amino acids 26–586 of human CD105 expressed in mouse myeloma cells.

Techniques: Control

Journal: Cells

Article Title: Microglial Phenotyping in Neurodegenerative Disease Brains: Identification of Reactive Microglia with an Antibody to Variant of CD105/Endoglin

doi: 10.3390/cells8070766

Figure Lengend Snippet: CD105/MAB1097-positive microglia associated with pathological structures. ( a ) panels A–B). Strongly CD105-positive microglia were mainly associated with Aβ plaques (panels A, B) but not with phospho-tau positive neurofibrillary tangles (C). CD105/RnD staining observed in p-tau positive neuritic plaque. Scale bars represent 50 μm. ( b ) Confocal colocalization of CD105 and Aβ. Colocalization of CD105/RnD (green) and Aβ (red) and merged image (yellow) demonstrated by confocal microscopy in AD case. The area delineated on Merge image encompasses a microglia with intracellular Aβ (yellow structures). Images are at the same magnification; scale bars represent 20 μm. ( c ) CD105-positive microglia containing neuromelanin. Panels A–D) Strongly positive CD105 positive structures (purple arrows) associated with neuromelanin particles (arrowheads) in the substantia nigra. Panel A, Control case; Panel B, ILBD case; and panel C and D, PD cases. Images are at the same magnification; scale bars represent 50 μm.

Article Snippet: CD105 antibodies from R&D Systems (Minneapolis, MN, USA) (MAB1097) or Abcam (Cambridge, MA, USA.) (AB221675) were mixed for 18 hours with recombinant CD105 protein (1097-EN, R&D Systems), which consists of amino acids 26–586 of human CD105 expressed in mouse myeloma cells.

Techniques: Staining, Confocal Microscopy, Control

Journal: Cells

Article Title: Microglial Phenotyping in Neurodegenerative Disease Brains: Identification of Reactive Microglia with an Antibody to Variant of CD105/Endoglin

doi: 10.3390/cells8070766

Figure Lengend Snippet: Levels of CD105–70 kDa polypeptide in in vitro-stimulated human brain microglia. Top panel: Bar chart showing mean levels of CD105 70 kDa polypeptide in stimulated human microglia. Microglia were stimulated with Aβ(42)(2 μM), poly IC (pIC) (25 μg/mL) or combination of both (pIC-Aβ) in triplicate for 24 hours and analyzed by Western blotting. Significant increase in 70 kDa in pIC-stimulated but not Aβ-stimulated microglia. Results normalized against values for β-actin. This is representative of 3-independent experiments with different microglia isolates. *** p < 0.001; ** p < 0.01. Lower panel: Western blot showing bands in treated microglia (MG). The same 70 kDa band was detected in sample of brain endothelial cells (BEC) isolated from same case. Blots were reprobed with antibody to β actin for normalization purposes.

Article Snippet: CD105 antibodies from R&D Systems (Minneapolis, MN, USA) (MAB1097) or Abcam (Cambridge, MA, USA.) (AB221675) were mixed for 18 hours with recombinant CD105 protein (1097-EN, R&D Systems), which consists of amino acids 26–586 of human CD105 expressed in mouse myeloma cells.

Techniques: In Vitro, Western Blot, Isolation