Figure 1

Live single cell TGF-β signalling promotes wound healing. A. MDA-MB-231 cells were treated without (i) or with (ii) TGF-β (5 ng/ml) then tracked for 10 h with images taken every 4 min. Slow moving cells (iii) and fast moving cells (iv) within the same cell population treated with TGF-β. B. MDA-MB-231 cells were infected with Ad.Cre-GFP and Ad.CAGA-Td-Tom virus, stimulated with TGF-β and imaged for both GFP and Td-Tom. C. MDA-MB-231 cells were infected with Ad.CMV-Td-Tom at varying MOI or D. Ad.CAGA-Td-Tom virus at a MOI of 2500. Following stimulation with ± TGF-β (5 ng/ml) for 24 h, cells were fixed, permeabilised and stained with DAPI. Percentage positivity was calculated by visualising Tomato expression (Red) compared to nuclear staining (blue). E. Wound Area at 0 and 24 h post wound after cells had been infected with Ad.CAGA-Td-Tom virus and stimulated with ± TGF-β (5 ng/ml). F. 24 h post wound, cells were fixed, permeabilised and nuclear stained as above and images were taken visualizing Smad3 active cells (red) and nuclear staining (blue). 42% (360 out of 867) cells were positive in the non-wound area versus 62% (175/279) cells in the wounded area. G. The relative pixel intensity of Smad3 activity was quantified (Average of 5 randomly chosen fields ± SD). These data are representative of at least 3 separate experiments (*P < 0.05).