Fig. 6

(A) 2D sections of an anterior bud showing cells (1, 2 and 3) engaged in a tether (red arrow) that eventually breaks and retracts. Scale bar, 10 μm.

(B) Two-dimensional (2D) and three-dimensional (3D) views of segmented cells showing membrane tether between cells 1,2 and 3 using side and transverse sections.

(C) 2D sections of a lateral bud showing cells (1, 2 and 3) engaged in a tether (red arrow) using mosaic membrane labelling in red and Myosin II (black) reporter. Scale bar, 10 μm.

(D) 2D section of an anterior bud showing accumulation of E-Cadherin (black) on the lateral membrane (blue arrows) and in the tether between cells 1 and 2 (red arrow)

(E) Side and transverse views of 2D sections showing tethered and unmembrane tether retraction with membrane labelling. The cell forming an unmembrane tether (top) is marked with an asterisk. The cells forming membrane tethers (bottom) are tracked with numbers 1 and 2. Adjacent membrane angles and adjacent cell distance are shown in blue and red before and after retraction respectively. Scale bar, 10 μm.

(F) Adjacent membrane angles (G) and cell distance (H) before and after retraction. ‘n’ denotes the number of cells. p values as labelled (Mann Whitney- U test).

(G)Left: 2D sections of lateral buds from controls (top) and HAase (bottom) injected embryos using Tg(βActin:membrane-Citrine) and Texas-red dextran in the periotic space. Membrane tethers (marked by white arrow) retract in control (top) but not in HAase treated embryo (bottom). Scale bar, 10 μm. Right: Individual data points and mean±s.d. of the duration of membrane tethers in control and HAase-treated embryos. n denotes the number of tethers. p values as labelled (Mann Whitney- U test).

(H) Left: 2D sections of anterior buds from controls (top) and DN E-Cadherin (bottom) expressing embryos using Tg(βActin:membrane-Citrine). Membrane tether (marked by white arrow) is present in control (top) but not in perturbed embryo (bottom). Scale bar, 10 μm. Right: Individual data points and mean±s.d. of the number of membrane tethers in control and DN E-cad mutant expressing embryos. n denotes the number of buds. p values as labelled (Mann Whitney- U test).