miR-155 increases lymphoma cell motility. (A) Raji, VAL, and MC116 cells were transfected with hsa-miR-155 or nontargeted control. Forty-eight hours later, the cells were used for SDF-1 chemotaxis assay performed in triplicate, as described in “Chemotaxis assays.” Data are mean ± SE. *Significant difference (P < .05). Western blot confirms down-regulation of HGAL protein levels in the hsa-miR-155–transfected cells. (B) Representative pictures demonstrating spontaneous movement paths of Raji cells, transfected with hsa-miR-155 or with nontargeted control. Forty-eight hours after transfection, cells were seeded on fibronectin-coated 6-well plates. Time lapse images were acquired every 2 minutes for 1 hour. Top portion: Motility tracks (in green) of selected cells (in red). Bottom portion: Motility tracks of 50 randomly selected cells over 1 hour, measured, and graphed using the Track Points. Each track was assigned a different random color. Timelapse images were acquired with a BD Pathway 855 HCS Bioimager (BD Biosciences) with an Olympus 10×/0.3NA objective lens and ORCA-ER CCD camera. Images were saved as 1344 × 1024 pixel TIFF images, opened in MetaMorph 5.07 (Universal Imaging–Molecular Devices) and generated the video and measured and graphed using Track Points. Data was exported to Microsoft Excel 2007 by dynamic data exchange. (C) Average velocity of 50 cells in each treatment group. *Significant difference (P < .05). Error bars represent SEM.