Journal:

Article Title: Angiopoietin-1 and Angiopoietin-2 Activate Trophoblast Tie-2 to Promote Growth and Migration during Placental Development

doi:

Figure Lengend Snippet: Effect of Ang-1 on trophoblast migration. Migration of ED77 was observed with a modified Boyden chamber. Cells (2 × 105) were seeded in the upper compartment in 0.5% FCS. A: Increasing concentrations of Ang-1 were placed in the lower compartment. B: Ang-1 was premixed with recombinant Tie-2-FC for 30 minutes before its addition to the lower compartment. C: The effect of Ang-2 on trophoblast migration was assessed in a fashion similar to that of the analysis of Ang-1 in B. Cells that migrated across the polycarbonate filters in response to the vehicle alone or to Ang-1/Ang-2 were counted after a 6-hour incubation. Chemotaxis in response to FCS (20%) (black column) was used as a positive control. Results are expressed as the mean (± SEM) ED77 counted per 10 fields (×200) in representative experiments performed in duplicate and quadruplicate, respectively. Five similar experiments were performed on different cell passages with similar results.

Article Snippet: A polyclonal rabbit anti-Tie-2 antibody (sc-324) was purchased from Autogen Bioclear (Stevenage, UK).

Techniques: Migration, Modification, Recombinant, Incubation, Chemotaxis Assay, Positive Control

Journal:

Article Title: Angiopoietin-1 and Angiopoietin-2 Activate Trophoblast Tie-2 to Promote Growth and Migration during Placental Development

doi:

Figure Lengend Snippet: Quantification of Tie-2 mRNA gestational human placenta. A: Identification of a 265-bp protected mRNA fragment for Tie-2 by RNase protection assay, using total placental RNAs from first (FT) (n = 4), second (ST) (n = 5), and third (TT) (n = 5) trimester, as well as term (n = 6). The abundance of mRNA for 28S rRNA is also shown for each sample for comparison of RNA amounts. B: Graphical representation of Tie-2 mRNA levels. The ratio of Tie-2 to 28S rRNA was calculated from the protected band intensities as assessed by laser densitometric analysis. Levels of Tie-2 increased with increasing gestation.

Article Snippet: A polyclonal rabbit anti-Tie-2 antibody (sc-324) was purchased from Autogen Bioclear (Stevenage, UK).

Techniques: Rnase Protection Assay

Journal:

Article Title: Angiopoietin-1 and Angiopoietin-2 Activate Trophoblast Tie-2 to Promote Growth and Migration during Placental Development

doi:

Figure Lengend Snippet: Immunohistochemical localization of Tie-2 receptor in the human placenta. Serial sections were incubated with a rabbit polyclonal anti-Tie-2 antibody (1:100). A and B: Intense immunostaining staining for Tie-2 receptor is demonstrated in the cytotrophoblast-syncytiotrophoblast bilayer of placental villi of term placenta. C: Intense immunostaining for Tie-2 is demonstrated in the cytotrophoblast and syncytiotrophoblast of first-trimester placenta. D: The endothelial cells of the placental blood vessels also display strong tie-2 immunostaining. The control is negative, with nonimmune antibody. Original magnification: A and B, ×100; C and D, ×400.

Article Snippet: A polyclonal rabbit anti-Tie-2 antibody (sc-324) was purchased from Autogen Bioclear (Stevenage, UK).

Techniques: Immunohistochemical staining, Incubation, Immunostaining, Staining

Journal:

Article Title: Angiopoietin-1 and Angiopoietin-2 Activate Trophoblast Tie-2 to Promote Growth and Migration during Placental Development

doi:

Figure Lengend Snippet: Identification of Tie-2 in a human first-trimester trophoblast cell line. A: Proteins were extracted from serum-starved HUVECs and trophoblast cells (ED27) and immunoprecipitated with anti-Tie-2 antibody (1 μg/100 μl) overnight at 4°C before Western blotting with anti-angiopoietin 1 (1:2000). Both endothelial (E) and trophoblast (T) extracts show a band of 208 kd corresponding to the Tie-2 receptor plus Angiopoietin 1. B: Cells were prepared and immunoprecipitated as in A before Western blotting with an anti-VEGF antibody (1:1000). No specific bands were observed in either endothelial or trophoblast cell extracts. The data shown are representative of three independent experiments.

Article Snippet: A polyclonal rabbit anti-Tie-2 antibody (sc-324) was purchased from Autogen Bioclear (Stevenage, UK).

Techniques: Immunoprecipitation, Western Blot