Figure 2.
Figure 2. MDCK cells exert forces on junction through myosin II–dependent contractility. (A–C) Time-lapse frames showing oscillations in junctional lengths of young and maturing monolayers. (D) Measurements in maturing monolayer showing junctional length deviations (purple arrows) from steady state (green arrowheads). The trace shown is from a single representative experiment out of 12 traces. (E) Plotting of 12 individual junctional lengths shows variations independent of the absolute lengths. (F) Measurements of mature monolayer showing junctional length (green arrowheads) with a flat steady state baseline. The trace shown is from a single representative experiment out of six traces. (G) Variations of junctional length decrease with maturation of cell monolayer. n = 6. (H) Blebbistatin inhibits oscillations of junctional length. (E and H) Means are represented by horizontal lines. (I and J) Time-lapse frames of mature monolayer showing transient increases in cell diameter (red and pink asterisks) from original (yellow and white asterisks) diameter. Pink and red lines show the diameters of cells used for analysis. (K) Measurements of cell diameter in maturing monolayer showing decrease in diameter (black arrow) away from steady state (green arrowheads). The trace shown is from a single representative experiment out of seven traces. (L) Measurements of cell diameter in mature monolayer showing fluctuations with transient decrease (black arrows) and increases (pink arrows). The trace shown is from a single representative experiment out of six traces. (M) Centripetal apical junctional tugging persists in mature monolayers. n = 40. (N) Blebbistatin inhibits centripetal apical junctional tugging. n = 30. (G, M, and N) Error bars are standard errors. (O) Contractile apparatus is associated with cell–cell junction in T84 epithelial cells. Yellow arrows point to dense bodies, characteristic of myosin II–containing sarcomeric contractile unit, with associating actin filaments attached to the cell junction (red arrows). Bars: (A–C) 2 µm; (I and J) 10 µm; (O) 100 µm.

MDCK cells exert forces on junction through myosin II–dependent contractility. (A–C) Time-lapse frames showing oscillations in junctional lengths of young and maturing monolayers. (D) Measurements in maturing monolayer showing junctional length deviations (purple arrows) from steady state (green arrowheads). The trace shown is from a single representative experiment out of 12 traces. (E) Plotting of 12 individual junctional lengths shows variations independent of the absolute lengths. (F) Measurements of mature monolayer showing junctional length (green arrowheads) with a flat steady state baseline. The trace shown is from a single representative experiment out of six traces. (G) Variations of junctional length decrease with maturation of cell monolayer. n = 6. (H) Blebbistatin inhibits oscillations of junctional length. (E and H) Means are represented by horizontal lines. (I and J) Time-lapse frames of mature monolayer showing transient increases in cell diameter (red and pink asterisks) from original (yellow and white asterisks) diameter. Pink and red lines show the diameters of cells used for analysis. (K) Measurements of cell diameter in maturing monolayer showing decrease in diameter (black arrow) away from steady state (green arrowheads). The trace shown is from a single representative experiment out of seven traces. (L) Measurements of cell diameter in mature monolayer showing fluctuations with transient decrease (black arrows) and increases (pink arrows). The trace shown is from a single representative experiment out of six traces. (M) Centripetal apical junctional tugging persists in mature monolayers. n = 40. (N) Blebbistatin inhibits centripetal apical junctional tugging. n = 30. (G, M, and N) Error bars are standard errors. (O) Contractile apparatus is associated with cell–cell junction in T84 epithelial cells. Yellow arrows point to dense bodies, characteristic of myosin II–containing sarcomeric contractile unit, with associating actin filaments attached to the cell junction (red arrows). Bars: (A–C) 2 µm; (I and J) 10 µm; (O) 100 µm.

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