Figure 1.
Figure 1. Properties of the “tailless” cadherin clusters. (a) Immunofluorescence microscopy of A431D cells expressing EcDnΔ (Δ) or its cis-interface-defective version, cis-EcDnΔ (Cis-Δ). The cells were stained with anti-Dendra antibody. Higher magnifications of the selected regions (indicated by dashed boxes) are shown in the insets. Note that cell–cell contacts exhibit numerous EcDnΔ clusters of different intensities. The individual clusters are not resolved at high level of mutant expression (arrow). The clusters became undetectable upon mutation of the cis-interface. Bars: (main images) 15 µm; (insets) 5 µm. (b) Time-lapse analyses of EcDnΔ clusters present in an area boxed in the left image. The sequence is assembled from Video 1. Some clusters, two of which are marked with blue and yellow circles, persist over the entire 1.5-s-long video. Others (red circles) are much more transient. Bars: (left) 10 µm; (right) 3 µm. (c) Time-lapse analyses of D1A-EcDnΔ clusters (from Video 2). Point mutation D1A increases stability of the cadherin trans-dimers. Note that this mutation significantly decreases cluster dynamics. Bar, 3 µm. (d) 1-s-long trajectories of the six fastest clusters in cells expressing EcDnΔ (Δ), D1A-EcDnΔ (D1A-Δ), and EcDnΔ-αABD (α). Two independent videos for each cell line were analyzed. Note that the EcDnΔ clusters (in Δ), although not moving in particular directions, covered distances of ∼0.2 µm. The D1A mutation or αABD notably restricted cluster motility. (e) Dendra photoconversion assay of different EcDnΔ mutants: EcDnΔ (Δ), D1A-EcDnΔ (D1A-Δ), EcDnΔ-αABD (Δ-αABD), and D1A-EcDnΔ-αABD (D1A-Δ-αABD). The graph shows changes in intensity of the red fluorescence over time (in seconds) in the entire laser-irradiated area (diameter = 0.25 µm; see Fig. S3). The error bars represent SDs (n = 30). (f) The lifetime of the arbitrary EcDnΔ (Δ) and D1A-EcDnΔ (D1A-Δ) clusters from two independent videos. Note that D1A-EcDnΔ clusters, in contrast to the parental EcDnΔ clusters, could typically be followed over the entire 2-s-long periods.

Properties of the “tailless” cadherin clusters. (a) Immunofluorescence microscopy of A431D cells expressing EcDnΔ (Δ) or its cis-interface-defective version, cis-EcDnΔ (Cis-Δ). The cells were stained with anti-Dendra antibody. Higher magnifications of the selected regions (indicated by dashed boxes) are shown in the insets. Note that cell–cell contacts exhibit numerous EcDnΔ clusters of different intensities. The individual clusters are not resolved at high level of mutant expression (arrow). The clusters became undetectable upon mutation of the cis-interface. Bars: (main images) 15 µm; (insets) 5 µm. (b) Time-lapse analyses of EcDnΔ clusters present in an area boxed in the left image. The sequence is assembled from Video 1. Some clusters, two of which are marked with blue and yellow circles, persist over the entire 1.5-s-long video. Others (red circles) are much more transient. Bars: (left) 10 µm; (right) 3 µm. (c) Time-lapse analyses of D1A-EcDnΔ clusters (from Video 2). Point mutation D1A increases stability of the cadherin trans-dimers. Note that this mutation significantly decreases cluster dynamics. Bar, 3 µm. (d) 1-s-long trajectories of the six fastest clusters in cells expressing EcDnΔ (Δ), D1A-EcDnΔ (D1A-Δ), and EcDnΔ-αABD (α). Two independent videos for each cell line were analyzed. Note that the EcDnΔ clusters (in Δ), although not moving in particular directions, covered distances of ∼0.2 µm. The D1A mutation or αABD notably restricted cluster motility. (e) Dendra photoconversion assay of different EcDnΔ mutants: EcDnΔ (Δ), D1A-EcDnΔ (D1A-Δ), EcDnΔ-αABD (Δ-αABD), and D1A-EcDnΔ-αABD (D1A-Δ-αABD). The graph shows changes in intensity of the red fluorescence over time (in seconds) in the entire laser-irradiated area (diameter = 0.25 µm; see Fig. S3). The error bars represent SDs (n = 30). (f) The lifetime of the arbitrary EcDnΔ (Δ) and D1A-EcDnΔ (D1A-Δ) clusters from two independent videos. Note that D1A-EcDnΔ clusters, in contrast to the parental EcDnΔ clusters, could typically be followed over the entire 2-s-long periods.

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