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Endothelial cells in REF and INV tumor sections were stained as described in “ ”. Microvessels are revealed by brown staining (magnification ×100) and endothelial cells were quantified (B). QRT-PCR was used to quantify VEGF , VEGFR2 , VEGFR1 and NRP-1 gene expression in INV and REF cells (C). VEGF (D) and NRP-1 (E) proteins were detected by ELISA tests and by western blotting, respectively. Each column represents a mean (± SD) of three independent experiments. * P <0.05. Gene functions from the GO Description provided by <t>NetAffx™</t> Analysis Center Affymetrix ( https://www.affymetrix.com/analysis/netaffx/index.affx )* Validated by QRT-PCR.
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Endothelial cells in REF and INV tumor sections were stained as described in “ ”. Microvessels are revealed by brown staining (magnification ×100) and endothelial cells were quantified (B). QRT-PCR was used to quantify VEGF , VEGFR2 , VEGFR1 and NRP-1 gene expression in INV and REF cells (C). VEGF (D) and NRP-1 (E) proteins were detected by ELISA tests and by western blotting, respectively. Each column represents a mean (± SD) of three independent experiments. * P <0.05. Gene functions from the GO Description provided by NetAffx™ Analysis Center Affymetrix ( https://www.affymetrix.com/analysis/netaffx/index.affx )* Validated by QRT-PCR.

Journal: PLoS ONE

Article Title: Invading Basement Membrane Matrix Is Sufficient for MDA-MB-231 Breast Cancer Cells to Develop a Stable In Vivo Metastatic Phenotype

doi: 10.1371/journal.pone.0023334

Figure Lengend Snippet: Endothelial cells in REF and INV tumor sections were stained as described in “ ”. Microvessels are revealed by brown staining (magnification ×100) and endothelial cells were quantified (B). QRT-PCR was used to quantify VEGF , VEGFR2 , VEGFR1 and NRP-1 gene expression in INV and REF cells (C). VEGF (D) and NRP-1 (E) proteins were detected by ELISA tests and by western blotting, respectively. Each column represents a mean (± SD) of three independent experiments. * P <0.05. Gene functions from the GO Description provided by NetAffx™ Analysis Center Affymetrix ( https://www.affymetrix.com/analysis/netaffx/index.affx )* Validated by QRT-PCR.

Article Snippet: Gene annotations and functions were assessed using NetAffx™ Analysis Center (Affymetrix, Santa Clara, CA, USA), Ingenuity pathway Analysis (IPA) software (Ingenuity SystemsInC.) and data mining.

Techniques: Staining, Quantitative RT-PCR, Expressing, Enzyme-linked Immunosorbent Assay, Western Blot