Figure 3

Cytoplasmic cyclin D1 controls the invasion and migration of MCL cells in vitro. (A) MCL cells were used to seed the top chamber of Transwell inserts. The inserts were transferred into wells containing medium supplemented with SDF1 and the cells were allowed to migrate for 4 h. The cells present in the bottom well were then counted. Three independent inserts were used per experiment and each experiment was performed three times. The mean numbers of migrating cells ± s.d. are plotted on the graph. ***p < 0.001 in Student’s t-test. (B) MCL cells were left untreated (controls) or treated with SDF1 (200 ng/ml for 4 h), then cytospun, fixed and permeabilized. F-actin was visualised with rhodamine-stained phalloidin by confocal microscopy (Fluoview FV 1000 confocal microscope and Fluoview Viewer software, Olympus). Nuclei were counterstained with DAPI (x180, magnification). (C) Cultured MCL cells were used to seed the ECM-coated 5 μm-pore membranes of Transwell inserts, in the top chamber, containing serum-free medium. The inserts were then placed in wells containing SDF1 as a chemoattractant and incubated for 24 h. The cells invading the lower chamber were counted by flow cytometry. The means ± s.d. of three independent experiments with triplicate samples are indicated on the graph. ***p < 0.001 in Student’s t-test.