Figure 7
TWIST1 mediates ARTEMIN (ARTN) stimulated attachment to, and migration through, an endothelial cell layer. (a) Forced expression of ARTN in BT549 and MDA-MB-231 cells promoted adhesion to an endothelial cell layer: Human microvascular endothelial cells1 (HMEC1) were grown to 100% confluence in 96-well plates and 5-chloromethylfluorescein diacetate (CFMDA) cell tracker green (5 μM) labelled BT549-ARTN and MDA-MB-231-ARTN cells ± TWIST1 siRNA were seeded on top for one hour, washed and counted under a fluorescence microscope. Bar, 78.4 μm. (b) Forced expression of ARTN in BT549 and MDA-MB-231 cells promoted in vitro trans-migration activity through an endothelial cell layer: CFMDA labelled cells were seeded on top of HMEC1 cells grown 100% confluence on transwell inserts. After 14 hours invaded cells were counted under fluorescence microscope. siRNA control-treated vector (VEC) cells presented as 100%. **, P < 0.01; ***, P < 0.001.