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. 2011 Oct 14;6(10):e26379. doi: 10.1371/journal.pone.0026379

Figure 4. NIH3T3 cells expressing Cx43dT-GFP show defects in wound closure and polarized cell movement.

Figure 4

(A–H) NIH3T3 cells transfected with plasmids expressing GFP, Cx43FL-GFP or Cx43dT-GFP were examined for wound closure 5 hrs after a wound scratch was made (n = 10, 10, and 8 scratches respectively). The rate of wound closure in the Cx43dT-GFP transfected cells (E, F) was reduced when compared with the GFP (A,B) or Cx43FL-GFP transfected cells (C,D). Quantitative assessment showed a significant decrease in wound closure rate in the Cx43dT-GFP expressing NIH3T3 cells at 4 and 5 hrs after wounding (G) (p<0.05). This was associated with a significant decrease in both the directionality and speed of wound closure (H). (I–L). Motion analysis with the tracing of individual cells at the wound edge showed a distinct polarized cell morphology in cells expressing GFP (n = 20 cells) (I) or Cx43FL-GFP (n = 20 cells) (J), with cytoplasmic protrusions (green) seen at the cell's leading edge facing the wound, and retracting cell processes (red) in the cell's trailing edge at the ipsilateral side. In contrast, in cells expressing the Cx43dT-GFP construct (n = 22 cells) (K), cell protrusions and retractions were not as distinctly polarized. This defect in cell polarity in cells expressing the Cx43dT-GFP construct was associated with an overall increase in cell protrusive activity (L). Data presented as mean ± SEM. Scale bars in (A–F) represent 100 µm. Scale bars in (G–I) represent 10 µm.