24 well plates with permeable membrane support inserts transwell  (Corning Life Sciences)

Journal: American Journal of Cancer Research

Article Title: ST3GAL2 knock-down decreases tumoral character of colorectal cancer cells in vitro and in vivo

doi:

Figure Lengend Snippet: Functional tests on HT29 cells. A. Cell cycle analysis by flow cytometry of ST3GAL2 KD and control cells. The percentages correspond to cells in G1 phase, S phase and G2/M phases. B. DNA fragmentation in HT29-shctrl and HT29-shST3GAL2 cells was quantified by ELISA. DNA degradation is expressed relative to control cells. Graph bars represent mean ± SD. C. HT29 cells were stained with Annexin V-FITC and PI, and analyzed by flow cytometry. Percentages represent living cells in lower left panel, cells in apoptosis in lower right panel, and dead cells in upper right panel. D. Migration assays were performed in free FBS medium to detect the cell migration ability of HT29 cells. The gap closure was measured every day and converted in closure percentage. Graph bars represent means of percentage closure ± SD. from three independent experiments. *P<0.05, ***P<0.001. E. Invasion tests: cells were placed in the upper chamber of a transwell insert, and matrigel-transwell assays were performed to detect every 24 h the cell invasion ability of HT29 cells by crystal violet staining of cells on the underside of the membrane. *P<0.05, **P<0.01, ***P<0.001.

Article Snippet: Briefly, the membrane (upper side) of transwell chamber (Transwell ® Permeable Supports 6.5 mm Insert, 24 Well Plate, 8.0 μm polycarbonate Membrane, Costar ® , Corning Incorporated, USA) was precoated with 0.5 mg·ml -1 matrigel (Corning ® Matrigel ® Basement Membrane Matrix, 354234, Discovery Labware, Inc., Two Oak Park, Bedford, USA), and cells (1×10 5 ) were placed in the top chamber.

Techniques: Functional Assay, Cell Cycle Assay, Flow Cytometry, Control, Enzyme-linked Immunosorbent Assay, Staining, Migration, Membrane