Fig. 5

Tandem fluorescent protein timers (tFTs) reveal changes in cadherin stabilisation at subcellular and tissue scales. (A) Images from a time-lapse movie showing the progressive front to rear loss of the apical Cdh2-GFP foci after colcemid treatment (right panels). A deposited neuromast maintaining Cdh2-GFP adherens junctions after colcemid treatment is shown on the left. (B) Box plots of the time required for the loss of the 1st, 2nd and 3rd rosettes after treatment with colcemid (n=12 pLLP, Friedman rank sum test, ^**P<0.001). (C) BAC cdh2:Cdh2-tFTs embryo illustrating the Cdh2 age differences between tissues. Inset, magnification of the squared area containing the pLLP. Maximal intensity projections. (D) Ratiometric images of cdh2:Cdh2-tFTs show a relative increase of the red/green ratio from the front to the back pLLP rosettes and deposited neuromasts. (E) Box plots of the distribution of the lifetime ratios (red/green) of apical clusters and subjacent membranes in the corresponding area demonstrating the increased lifetime of Cdh2 in the clusters compared with membranes (n=175, paired t-test, ^***P<0.001). (F) Mean and s.e.m. of the age difference between clusters and membranes (red/green ratios of clusters minus membrane) normalised on the pLLP leading most cluster (rosette1) showing a gradual stabilisation of Cdh2 in the clusters compared with membrane as rosettes mature into neuromasts (n=35 pLLP, Friedman rank sum test, ^***P<0.001). NM, neuromast. Scale bars: 200 µm in C; 20 µm in A,D.