All mutants exhibit sensitivity to prolonged Ca<sup>2+</sup>-chelation.

<p>(<b>A</b>) Example images for WT and all mutants before and 1 h after BAPTA addition. Scale bar = 10 µm. (<b>B</b>) Quantification of junction disassembly. Mean fluorescence intensity of multiple junctions from the different genotypes was measured over time. The drop in mean fluorescence intensity is specific to the addition of BAPTA (blue), indicated by the arrow, as application of a vehicle (green) did not lead to a decrease in junction intensity. (<b>C</b>) Comparison of junction disassembly between different mutants and WT. Although the time course and degree of disassembly differed for the various mutants, with W2A junctions displaying the highest degree of disassembly; on a longer timescale, none of them were completely Ca<sup>2+</sup>-insensitive. N-cadherin-R14E, which appeared Ca<sup>2+</sup>-insensitive on a short timescale (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0081517#pone-0081517-g003" target="_blank">Fig. 3 D</a>) also showed a drop in junction intensity 10 min after BAPTA application. Error bars indicate SEM. n(WT) = 84, n(W2A) = 56, n(R14E) = 84, n(V81D/V174D) = 52 junctions. </p>

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