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Fig. 1. Serum starvation of NIH3T3 cells results in failure of trailing-edge retraction. (A) Length of the trailing edge and protrusion rates in NIH3T3 cells stably expressing βactin-mRFP cultured without serum or without serum but plus 2 µM LPA. For trailing-edge measurement, cells were filmed for 6 hours immediately after serum removal with images acquired at 5-minute intervals, using phase-contrast microscopy. Images acquired at the end of the sixth hour were quantified are shown. Data represent the analysis of 60 cells from two different experiments. For protrusion of the lamellipodia, cells were starved for 6 hours and at the end of sixth hour, images of βactin-mRFP in protruding lamellipodia were obtained for 20 minutes at 20-second intervals. The data represent analysis of six protruding lamellipodia per experimental condition. (B) Phase-contrast images and quantification of cell elongation of serum-starved cells, serum-starved cells treated with 2 µM LPA or 100 nM PDGF. Data represent the analysis of 50-72 cells per condition from two different experiments. (C) Kinetics of changes in cell elongation following LPA stimulation of serum-starved fibroblasts.
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