siRNA against PPL reduces cell migration in D562 cells. D562 cells were transfected with control and PPL siRNA. (A), (B) Quantification of wound closure in control and PPL siRNA. Each cell was grown to confluence and then wounded by scraping. For each wound, the width of the wound at 0 h was designated as 100%, and the subsequent timepoints in the graph show the relative width of the open wound. Values represent the mean ± S.E. from 3 separate experiments. Asterisks indicate a p value less than 0.01 based on Student's t-test. Bar: 200 μm. (C), (D) Paths of 2 transfected cells from a time-lapse recording. D562 cells were transfected with either control or PPL siRNA. After 24 h, cells were seeded on glass-bottomed dishes and incubated for another 24 h. Time-lapse recordings were made every 5 min for 24 h. Migration distance was measured using 10 control and 10 PPL siRNA-transfected cells. Bar: 100 μm. Asterisks indicate a p value less than 0.05 based on Student's t-test. (E) Downregulation of PPL resulted in reduced pAkt expression. The expression of PPL and β-actin was analyzed by western blotting with total cell extracts from transient transfections with control siRNA (GL2) and PPL siRNA 1 and 2.
Tonoike et al. BMC Cell Biology 2011 12:41 doi:10.1186/1471-2121-12-41