294150.fig.001
Figure 1: Adenoviral-mediated overexpression of BMP-2 and its effect upon cellular motility in both uni- and multicell populations. (a) GFP expression of adenoviral transfected rat VSMCs assessed by confocal microscopy. Infection efficiency exceeds 90%. (b) Rat VSMCs migration traced by time-lapse video microscopy. Sixteen representative paths for each treatment originated from a common point, in stellate fashion. Faster migration produced larger stars. Cellular motility was recorded on indicated substrates for 6 hours. (c) Movement assay: mean velocity determined by time-lapse recording via ImageJ software. Overexpression of BMP-2 approximately doubled migratory capacity. (d) Boyden chamber assay revealed increased cellular migration in adenoviral-transfected cells overexpressing BMP-2 compared to vector-only cells. (e) Quantification of migration of BMP-2 overexpressing transfected cells; values represent the mean ± SD of independent experiments. (f) Wounding assay, assessing motility, and spread potential of a cellular population. VSMCs cells were virally-transfected to overexpress BMP-2. Both at the time of cellular culture incision and 48 hours after, the state of wound closure was assessed by phase-contrast microscopy. Arrows indicate distance from incision edges. *Student’s -test: versus control.