Fig. 2: PTEN controls first-row cell-cell cohesion.

a, d Normal (a) and super resolution (d) immunofluorescence images of actin filaments (phalloidin), cell-cell junctions (N-cadherin) in siCTL and siPTEN#1 migrating astrocytes. Black arrow: direction of migration. Boxed regions are zoomed in the panels below to highlight the presence of interjunctional transverse actin arcs (ITA) mostly in siCTL cells (black arrowheads). The micrographs in (a) are representative of the data shown in (b, c). The images in (d) are representative of six independent samples. b Angular distribution of actin filaments in front row siCTL and siPTEN#1 cells (n = 300). 0° is perpendicular to the direction of migration. Graph represents Kernel density estimates of probability density function. Dashed lines show quartiles. c, e Proportion of front row siCTL and siPTEN#1 cells with (e, n = 200) or without (c, n = 300) wt- and mutated-PTEN rescue, that are connected with ITA. f Fluorescence images of CCV ECs expressing LifeAct-GFP in CTL and ptenb morphant fish. White arrowheads point at ITA. g Percentage of front row CCV EC cells connected by ITA in MoCTL (n = 36 cells) and MoPTENb (n = 29 cells) fish larvae. Box extends from the 25th to 75th percentiles and the line in the middle of the box is plotted at the median. The whiskers delineate all data points from minimum to maximum. h Immunofluorescence image of N-cadherin in migrating astrocytes, where the linearity index of front row lateral cell-cell junction is defined. i Junctional linearity index in siCTL (n = 70) and siPTEN#1 (n = 65) cells, 8 h after migration. j Junctional linearity index in MoCTL (n = 12) and MoPTENb (n = 16) front row cells, 100 min after migration. k Schemes summarising PTEN loss phenotype in front row cells during collective cell migration. Scale bars: 10 µm. Number of independent experiments = 2 (e, i), =3 (b, c, j). Statistical tests: Kolmogorov-Smirnov (b), two-tailed paired t-test (c, g), two-tailed Mann–Whitney (i), two-tailed unpaired Student t test (j). p values, along with effect size coefficients are given on the graphs. Error bars represent SD. Source data are provided as a Source Data file.