Samples from each group were fixed with 4% paraformaldehyde for 24 h and subsequently incubated with 0.5% Triton-X for 10 min. Bovine serum albumin (1%) was used to block the cells for 1 h. Samples were dehydrated in sections after paraffin embedding. After blocking with 5% goat serum (Life Technologies, New York, NY, United States) for 1 h at 20 °C, the specimens were then treated with rabbit anti-CD31 (Cell Signaling Technology, Boston, United States), anti-VE-cadherin (Cell Signaling Technology, Boston, United States), FGF2 (Ab208687-40, Abcame, Shanghai, United States), anti-FOXO1 (Cell Signaling Technology, Boston, United States), or anti-p-AKT (Cell Signaling Technology, Boston, United States) antibodies for 12 h at 4 °C. Alexa Fluor 594-conjugated goat anti-rabbit secondary antibody (GB25303; Sevicebio, Wuhan, China) was added, and the cells were incubated for 2 h at room temperature. The nuclei were stained with 4’,6-diamidino-2-phenylindole. CD31, VE-cadherin, FGF2, FOXO1, and p-AKT expression was observed under a fluorescence microscope (Nikon Eclipse, Tokyo, Japan). Statistical analysis was performed using GraphPad Prism 7 Software (San Diego, CA, United States).
Ve cadherin
Manufactured by Nikon
Sourced in Japan
VE-cadherin is a cell adhesion molecule that plays a crucial role in the formation and maintenance of endothelial cell-cell junctions. It is essential for the regulation of vascular permeability and the integrity of blood vessels.
Automatically generated - may contain errors
Lab products found in correlation
Goat serum, by Thermo Fisher Scientific (1 mentions)
Anti p akt, by Cell Signaling Technology (1 mentions)
Anti ve cadherin, by Cell Signaling Technology (1 mentions)
Anti foxo1, by Cell Signaling Technology (1 mentions)
Anti cd31, by Cell Signaling Technology (1 mentions)
Nucblue, by Thermo Fisher Scientific (1 mentions)
E cadherin, by Nikon (1 mentions)
2 protocols using ve cadherin
1
Immunofluorescence Staining of Vascular Markers
2
Microscopic Imaging of Cell-Cell Junctions
Check if the same lab product or an alternative is used in the 5 most similar protocols
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We used the live nuclear dye NucBlue (ThermoFisher; a Hoechst 33342 derivative) with a 30 min incubation for nuclear labeling on standard MDCK, HUVEC, and MDA-MB-231 tissues and imaged with a DAPI filter set. For MDCK data collected for pre- and post-contact inhibition experiments, nuclear labels were reproduced using a convolutional neural network trained to reconstruct nuclei features from 4x phase contrast images of cells. Complete documentation including code and trained network weights for this tool may be referenced in [39 (link)]. Media was swapped and silicone microwell stencil was removed prior to imaging. Cadherin imaging was performed using conventional epifluorescence microscopy on a Nikon Ti2 equipped with a YFP filter set (HUVEC VE-cadherin) and an RFP filter set (MDCK E-cadherin).
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