JCS00600 Movies

Files in this Data Supplement:

• Movie 1 - Migration of BAE cells in serum-free DMEM alone. BAE cells were loaded onto the Dunn chamber in serum-free media alone. Under these conditions, BAE cells show minimal migration and no directional orientation. Images were taken every 2 minutes for 6 hours and 40 minutes (200 frames), and videos were created using Metamorph software at 30 frames per second. Video correlates with migration response depicted in Table 1 and Fig. 9A.
• Movie 2 - Hep I-induced BAE cell chemokinesis. BAE cells were loaded onto the Dunn chamber in the presence of 100 nM hep I in both the inner and outer well, so as to stimulate focal adhesion disassembly but not present the cells with a chemical gradient. Hep I induces an increase in chemokinetic migration in the BAE cells, characterized by an increase in cell speed, the percent of migrating cells and the total cellular displacement. Directionality of migration is not significantly different from serum-free media alone, and cells display no preference for migration orientation. Images were taken every 2 minutes for 6 hours and 40 minutes (200 frames), and videos were created using Metamorph software at 30 frames per second. Video correlates with migration response depicted in Table 1 and Fig. 9B.
• Movie 3 - LRP-knockout MEFs are migration deficient. LRP-knockout MEFs were loaded onto the Dunn chamber in the presence of serum-free DMEM, and migration measured over a 6 hour and 40 minute time span. LRP-knockout fibroblasts show no migration with little to no lamellipodia extension, suggesting a defect in lamellipodia formation underlies the LRP-knockout migratory phenotype. Images were taken every 2 minutes for 6 hours and 40 minutes (200 frames), and videos were created using Metamorph software at 30 frames per second. Video correlates with migration response depicted in Fig. 7.
• Movie 4 - Endothelial cells display poor chemotaxis to an aFGF gradient. BAE cells were loaded onto the Dunn chamber in the presence of an aFGF gradient, with highest concentrations of aFGF (67 pM) on the right side of the screen. BAE cells display little migration orientation in response to the aFGF gradient. Images were taken every 2 minutes for 6 hours and 40 minutes (200 frames), and videos were created using Metamorph software at 30 frames per second. Video correlates with migration response depicted in Fig. 9D.
• Movie 5 - bFGF stimulates a potent chemotactic response in BAE cells. BAE cells were loaded onto the Dunn chamber in the presence of a bFGF gradient, with highest concentrations (61 pM) on the left side of the screen. BAE cells show a strong migration orientation towards the bFGF gradient. Images were taken every 2 minutes for 6 hours and 40 minutes (200 frames), and videos were created using Metamorph software at 30 frames per second. Video correlates with migration response depicted in Fig. 9G.
• Movie 6 - Enhanced aFGF-induced chemotaxis in the presence of hep I. BAE cells were loaded onto the Dunn chamber in the presence of a constant concentration of hep I (100 nM) and an aFGF gradient with highest concentrations (67 pM) on the right side of the screen. Most cells migrate towards the aFGF gradient, suggesting that hep I enhances the ability of aFGF to orient BAE cell migration. Images were taken every 2 minutes for 6 hours and 40 minutes (200 frames), and videos were created using Metamorph software at 30 frames per second. Video correlates with migration response depicted in Fig. 9E.
• Movie 7 - Hep I abrogates bFGF-induced chemotaxis. BAE cells were loaded onto the Dunn chamber in the presence of a constant concentration of hep I (100 nM) and a bFGF gradient with highest concentrations of bFGF (61 pM) on the left side of the screen. Whereas hep I and bFGF stimulate a potent chemokinetic response, BAE cells lose the ability to orient their migration in response to a bFGF gradient. Images were taken every 2 minutes for 6 hours and 40 minutes (200 frames), and videos were created using Metamorph software at 30 frames per second. Video correlates with migration response depicted in Fig. 9H.