Figure 9. The PHLPP-mediated regulation of cell migration relies on its ability to control integrin expression.

A. Stable control and PHLPP knockdown ASPC-1 cells were subjected to Transwell migration analysis using HGF as the chemoattractant. To block integrin activation, ASPC-1 cell suspensions were incubated with blocking antibodies of β1 or β4 integrin in serum-free media containing 1% BSA for 20 minutes at 4°C. The cells were then seeded onto the collagen coated Transwells in the presence of the blocking antibodies and allowed to migrate for 4 hours at 37°C. Each experiment was done in duplicates and three independent experiments were averaged and expressed as mean ± SEM (* p<0.01 by two-sample t-tests compared to the control cells). B. Cells as treated as above with blocking antibodies were monitored for single cell motility using time-lapse imaging. The cells were allowed to migrate in response to HGF for 6 hours. The average velocities of 12 cells from each group are shown. Data represent mean ± SEM (n = 12 cells/line, * p<0.05 by two-sample t-tests).