Modifications in migration and spreading of cells by Borealin mutants. (A) Tracking of GFP-Nthy transfected with Borealin-wt, Borealin-114, Borealin-148 and Borealin-177 during 8 h after wound healing assay on time-lapse video microscopy as described in "Materials and Methods." (B) The length of migration in um was measured by quantifying the total distance that the positively transfected cells (GFP) moved from the edge of the wound toward the center of the wound in 8 h. Mean of the length of the migration from four independent experiments. *P = 0.04, 0.0036, 0.0202 respectively for Nthy with Borealin-114, Borealin-148 and Borealin-148 in comparison with Nthy with Borealin-wt, calculated by t test. (C) Analysis of spreading by immunofluorescence. Staining of Borealin-Flag (in green) and filamentous actin (in red) showed decreased spreading of Borealin-114, Borealin-148, Borealin-177 Nthy compared to Borealin-wt Nthy (NT, no transfected; mock, empty vectors). (D) The percentage of spread and unspread Nthy was estimated by counting Nthy for each cell line (light gray, unspread; medium gray, weakly spread; dark gray, strongly spread). The graph represents the mean five independent experiments. *P< 0.05, P < 0.01 calculated by t test.