Figure 4.

PACSIN 2 repression of cellular migration is dependent on cyclin D1a. (A) Cyclin D1^−/− MEFs were transduced with viral expression vector (MSCV-IRES-GFP) encoding cyclin D1a, cyclin D1b or GFP control. (B) Cells were transiently transfected with siPACSIN 2. Western blot was performed with an anti-PACSIN 2 antibody to show a reduction of endogenous PACSIN 2. (C) Transwell migration assays were conducted in triplicate. Cells that migrated were stained and counted. Five fields in each of triplicate wells were randomly recorded. (D and E) The time-lapse video-microscopy was conducted in cyclin D1^+/+ and cyclin D1^−/− MEFs transfected with a PACSIN 2 expression or control vector. Vector expressing GFP was co-transfected. The GFP-positive cells were further analyzed for cell migration velocity. Student t-test was used for quantitative analysis of cell migration.