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Figure 2


Fig. 2. Time course of epithelial disintegration and SRF induction. (A) Scheme of the reporter plasmid for life imaging. Three SRF-binding sites were inserted in front of an EGFP expression cassette containing a PEST domain for fast proteasomal degradation. (B) Time-lapse images of living cells harbouring the EGFP reporter. Confluent monolayers of stably transfected MDCK cells were microscopically analysed to monitor EGFP expression (upper panel) and cell shape changes (lower panel) upon Ca2+ switch to 0.02 mM for the indicated times. Mock indicates medium exchange to normal (1.8 mM) Ca2+ for 7 hours to exclude any potential alterations by addition of fresh medium or shear forces. (C) Induction profile showing the time course of EGFP expression in B. The arbitrary fluorescence intensity was calculated by subtracting the fluorescence at each time point in the mock experiment from the equivalent fluorescence value upon Ca2+ withdrawal.





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