Cell migration and invasion rates were determined as described in previously published methods (31 –33 (link)) in HOS, hFOB 1.19 and hFOB 1.19 DAB2IP shRNA cells. To study migration, 2×104 cells were seeded into each well of a 96-well plate. When cells had reached 80–90% confluence, a wound was generated using the Cellplayer 96-well Woundmaker (Essen BioScience, Inc., Ann Arbor, MI, USA). The images of cells were automatically acquired every 2 h for 24 h using the Incucyte LiveCell Imaging system (Essen BioScience, Inc.).
To study invasion, a scratch was made by using the Cellplayer 96-well Woundmaker, when cells had reached 80–90% confluence. Next, the wound was covered with 50 µl of Matrigel solution (1 mg/ml Matrigel in PBS) that was allowed to set for 1 h at 37°C. Appropriate growth medium (100 µl) was added and representative images were recorded at 2 h intervals for the duration of the experiment (24 h). All images were processed using the IncuCyte™ software package version 20151.1 (Essen BioScience, Inc.) to measure cell migration and invasion by obtaining the relative wound density.
To study invasion, a scratch was made by using the Cellplayer 96-well Woundmaker, when cells had reached 80–90% confluence. Next, the wound was covered with 50 µl of Matrigel solution (1 mg/ml Matrigel in PBS) that was allowed to set for 1 h at 37°C. Appropriate growth medium (100 µl) was added and representative images were recorded at 2 h intervals for the duration of the experiment (24 h). All images were processed using the IncuCyte™ software package version 20151.1 (Essen BioScience, Inc.) to measure cell migration and invasion by obtaining the relative wound density.