Figure 7
Effect of hypoxic simulation on the migration and invasion of breast cancer cells. Panel A: YS1.2 and pII cells were treated with either vehicle (control), or with or CoCl2 or DFO (100 µM) for 24 h. Cells were loaded into wells formed in an agarose layer in 6 well plates. The total number of cells migrating/penetrating into the agarose in both lateral directions were manually counted using an inverted microscope as described in Methods. Histobars represent mean ± SEM of 3 independent determinations. Panel B: YS1.2 and pII cells pre-treated with vehicle (C, open bar), CoCl2 or DFO (100 µM, hatched bars) were loaded into the top chamber of a Cultrex dish. Cells invading through the BME layer into the lower chamber containing either serum free media (SFM, solid bar) or media containing 40% serum (used as chemoattractant) were measured as described in Methods and represented here as relative fluorescence units. Histobars represents mean ± SEM of 6 independent determinations. Panel C: Snapshots from time-lapse photography of pII and EII cells cultured for 48 h. Cells were seeded separately into adjacent chambers of an Ibidi chamber slide with a space between them. After 24 h the scaffold was removed leaving a gap between the two monolayers. The time-lapse photography showed that only the pII cells moved towards the EII cell monolayer and over 27 h completely closed the gap. (10x magnification). A schematic diagram explaining each experimental setup is shown on the top of each panel.
