File:Survival-advantages-conferred-to-colon-cancer-cells-by-E-selectin-induced-activation-of-the-PI3K-1471-2407-11-285-S3.ogv

English: An irrelevant control siRNA does not influence the adhesion of colon cancer cells to E-selectin-expressing endothelial cells in a laminar flow chamber. HUVECs were trypsinized and grown for 24 hrs on gelatin-coated slides. These endothelial cells were treated with 20 ng/ml IL-1β for 4 h to induce the expression of E-selectin. The endothelial cell cultures were then placed in a laminar flow chamber under a shear stress of 1 dyne/cm2. Live HT29 cells were transfected by electroporation with control siRNA purchased from Qiagen. Tumor cells in suspension (2 × 106 per assay) were labeled for 30 min with Calcein AM and washed twice with M199 medium before being added in the medium circulating over endothelial cells. Videos were taken directly using a camera mounted on a TE2000 fluorescence microscope at ×20 magnification to follow the adhesion of HT-29 cells to HUVECs.