Figure S1.
Junction morphology and dynamics in claudin/JAM-A KO cells. (A) Variation in the apical cell area. Apical cell area was more variable in claudin/JAM-A KO cells. Data represent the ratio between the SD of the apical area to the mean apical area and are presented as mean ± SD (n = 9). **P < 0.005, t test. (B) The magnitude of stretching during spontaneous cell stretching events. MDCK II cells were exposed to larger degree of stretching. Data represent the magnitude of stretching of individual spontaneous cell stretching events and are presented as mean ± SD (n = 21). **P < 0.005, t test. (C) Duration of the spontaneous stretching events. The stretching events in MDCK II cells significantly persisted longer. Data represent the duration of individual spontaneous cell stretching events and are presented as mean ± SD (n = 21). **P < 0.005, t test. (D) The maximum speed of stretching during individual spontaneous cell stretching events. The speed of stretching did not significantly differ between MDCK II and claudin/JAM-A KO cells. Data represent the maximum stretching speed of individual spontaneous cell stretching events and are presented as mean ± SD (n = 21). n.s.: not significant, t test. (E) Scatter plot of the cell stretching speed and the ZO-1 intensity changes in multiple cell stretching events in MDCK II cells (white circles) and claudin/JAM-A KO cells (blue triangles). White circles: stretching in MDCK II cells, open blue triangles: stretching in claudin/JAM-A KO cells without junction breakage, and closed blue triangles: stretching in claudin/JAM-A KO cells with junction breakage. Junction breakage was observed along a wide range of stretching speeds. (F) Snapshots from a time-lapse movie of claudin/JAM-A KO cells expressing ZO-1-GFP that underwent junction breakage (arrows) despite the small magnitude of cell stretching. The cell (asterisk) underwent repetitive sequential cell stretching, and the junctions fractured shortly after the final stretching event began. (G) Quantification of the cell circumference (black) and ZO-1-GFP intensity (green) during the cell stretching event in claudin/JAM-A KO cells shown in F. The stretching events are highlighted by the shaded area. The cell experienced repetitive sequential stretching events and the junction fractured shortly after the final stretching event started. (H) Strain map showing the length change of individual cell junctions immediately before the junction breakage upon spontaneous cell stretching. Junctional strain correlated with the sites of junction breakage (arrows). (I) Strain map showing the length change of individual cell junctions immediately before the junction breakage upon cell division. Junction breakage (arrows) did not correlate with junctional strain. Panels H and I are cropped from the same movie (Video 2) shown in Fig. 2, C and D. Scale bars: 10 μm.

Junction morphology and dynamics in claudin/JAM-A KO cells. (A) Variation in the apical cell area. Apical cell area was more variable in claudin/JAM-A KO cells. Data represent the ratio between the SD of the apical area to the mean apical area and are presented as mean ± SD (n = 9). **P < 0.005, t test. (B) The magnitude of stretching during spontaneous cell stretching events. MDCK II cells were exposed to larger degree of stretching. Data represent the magnitude of stretching of individual spontaneous cell stretching events and are presented as mean ± SD (n = 21). **P < 0.005, t test. (C) Duration of the spontaneous stretching events. The stretching events in MDCK II cells significantly persisted longer. Data represent the duration of individual spontaneous cell stretching events and are presented as mean ± SD (n = 21). **P < 0.005, t test. (D) The maximum speed of stretching during individual spontaneous cell stretching events. The speed of stretching did not significantly differ between MDCK II and claudin/JAM-A KO cells. Data represent the maximum stretching speed of individual spontaneous cell stretching events and are presented as mean ± SD (n = 21). n.s.: not significant, t test. (E) Scatter plot of the cell stretching speed and the ZO-1 intensity changes in multiple cell stretching events in MDCK II cells (white circles) and claudin/JAM-A KO cells (blue triangles). White circles: stretching in MDCK II cells, open blue triangles: stretching in claudin/JAM-A KO cells without junction breakage, and closed blue triangles: stretching in claudin/JAM-A KO cells with junction breakage. Junction breakage was observed along a wide range of stretching speeds. (F) Snapshots from a time-lapse movie of claudin/JAM-A KO cells expressing ZO-1-GFP that underwent junction breakage (arrows) despite the small magnitude of cell stretching. The cell (asterisk) underwent repetitive sequential cell stretching, and the junctions fractured shortly after the final stretching event began. (G) Quantification of the cell circumference (black) and ZO-1-GFP intensity (green) during the cell stretching event in claudin/JAM-A KO cells shown in F. The stretching events are highlighted by the shaded area. The cell experienced repetitive sequential stretching events and the junction fractured shortly after the final stretching event started. (H) Strain map showing the length change of individual cell junctions immediately before the junction breakage upon spontaneous cell stretching. Junctional strain correlated with the sites of junction breakage (arrows). (I) Strain map showing the length change of individual cell junctions immediately before the junction breakage upon cell division. Junction breakage (arrows) did not correlate with junctional strain. Panels H and I are cropped from the same movie (Video 2) shown in Fig. 2, C and D. Scale bars: 10 μm.

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