Journal: Molecular Cancer Therapeutics

Article Title: A Human Monoclonal Anti-ANG2 Antibody Leads to Broad Antitumor Activity in Combination with VEGF Inhibitors and Chemotherapy Agents in Preclinical Models

doi: 10.1158/1535-7163.mct-09-0554

Figure Lengend Snippet: Figure 1. Antibody 3.19.3 binds human Ang2 and blocks Tie2-mediated phosphorylation and downstream signaling. A, the ability of antibody 3.19.3 to selectively bind Ang2 in a competitive Tie2-Fc ELISA assay was determined with an IC50 value of 506 pmol/L, whereas Ang1 binding was essentially undetectable. B, inhibition of Ang2 binding to human Tie2 in a cell-based HEK293 assay. Antibody 3.19.3 was tested for the ability to block Ang2 interactions with Tie2 expressed on the cell surface using biotinylated human Ang1 or Ang2. Percentage of inhibition of binding was calculated as a fraction of the maximum signal obtained without antibody and suggested EC50 values of 150 pmol/L affinity to Ang2, whereas the Ang1 binding did not reach saturation up to concentrations of 1 μmol/L antibody. C, inhibition of Ang2-induced Tie2 phosphorylation in HEK293/Tie2 cells. IgG control antibody (lanes 1, 2) was compared alongside 3.19.3 (lanes 3, 4) for the ability to block recombinant Ang2 (1 μg/mL)–mediated Tie2 phosphorlylation at 2:1 molar ratios (lanes 1, 3) or 1:2 molar ratios (lanes 2, 4) corresponding to 1.35 μg/mL or 5.4 μg/mL antibody, respectively. Controls for maximal Ang2 induction are shown (lane 5) or no treatment (lane 6), indicating baseline pTie2 activity in this assay.

Article Snippet: Cells were harvested and lysed, and 0.8 mg total protein was immunoprecipitated using an anti-Tie2 antibody (MAB313, R&D Systems).

Techniques: Phospho-proteomics, Enzyme-linked Immunosorbent Assay, Binding Assay, Inhibition, Blocking Assay, Control, Recombinant, Activity Assay

Journal: The American journal of pathology

Article Title: Adenovirus-mediated gene transfer of TGF-β1 to the renal glomeruli leads to proteinuria.

doi: 10.1016/j.ajpath.2011.11.023

Figure Lengend Snippet: Figure 7. A: Immortalized podocytes were treated with rTGF-1. B: TGF-1 significantly inhibited nephrin and synaptopodin expression in a dose-response manner, while increasing the expression of the angiopoietin receptor Tie2. *P 0.001, †P 0.01, and ‡P 0.05 versus untreated cells. C and D: Podocin was also significantly down-regulated by TGF-1. Untreated mouse cardiac endothelial cells (MCEC) were used as a positive control for Tie2 expression. E: MCECs were grown to confluence and exposed to AdDL, AdTGF-1, or no adenovirus (Control). Conditioned media were then added to podocyte culture. F: Nephrin expression was significantly down-regulated by MCEC media condition with AdTGF-1. G: Synaptopodin expression was not significantly affected.

Article Snippet: Cells were also treated with rTGF- 1 (5 ng/mL) and anti-human Ang1, Ang2 antibody, or a blocking anti-Tie2 antibody (R&D Systems).

Techniques: Expressing, Positive Control, Control

Journal: Microscopy and Microanalysis

Article Title: Immunofluorescent detection of Tie1 in the endothelium of the Rat and Human corpus cavernosum during aging

doi: 10.1017/s1431927613000810

Figure Lengend Snippet: Fig. 1 – Dual immunolabeling of Tie1 (green) and PECAM-1 (red) (Fig. 1a - exposure time 226 ms), and α- actin (red) (Fig. 1b – exposure time 178 ms) in corpus cavernosum of Rat aged 24 months. Note co-localization in the endothelium (white arrow) absent in smooth muscle layer (Fig. 1b). Separate labelling of green and red channels (Tie1 and PECAM-1, respectively) shown in Fig. 1A.

Article Snippet: Paraffin embedded sections were deparaffinized, rehydrated and submitted to epitope retrieval with heated Tris-EDTA pH 9.0 buffer before dual-immunolabeling of Tie1 (rabbit anti-Tie1 antibody -abcam) and specific markers of endothelium and smooth muscle cell (goat- anti-PECAM-1 and mouse anti-α-actin antibodies, Santa Cruz Biotechnology Inc. and Millipore, respectively).

Techniques: Immunolabeling

Journal: Microscopy and Microanalysis

Article Title: Immunofluorescent detection of Tie1 in the endothelium of the Rat and Human corpus cavernosum during aging

doi: 10.1017/s1431927613000810

Figure Lengend Snippet: Fig. 2 – Dual immunolabeling of Tie1 (green) and α-actin (red) in corpus cavernosum of young (Fig. 2a - exposure time 191 ms) and aged (Fig. 2b - exposure time 109 ms) Human. In both conditions, Tie 1 did not label smooth muscle cell, but its expression in the endothelium of the old individual appeared up-regulated (white arrow).

Article Snippet: Paraffin embedded sections were deparaffinized, rehydrated and submitted to epitope retrieval with heated Tris-EDTA pH 9.0 buffer before dual-immunolabeling of Tie1 (rabbit anti-Tie1 antibody -abcam) and specific markers of endothelium and smooth muscle cell (goat- anti-PECAM-1 and mouse anti-α-actin antibodies, Santa Cruz Biotechnology Inc. and Millipore, respectively).

Techniques: Immunolabeling, Expressing