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Finally, preferential interaction of DCL cells to EVL cell borders was described as a gaussian potential centred along the EVL cell border (Supplementary Fig. 17).

Data from a given rat were used only if both cannulae were within the medial NAc shell62 (including the shell/core or shell/olfactory tubercle border) (Supplementary Fig. 5); based on these criteria, two rats were excluded from the final analysis.

(h) In embryos expressing lifeact-GFP and overexpressing Rac1-T17N, Dcellslls barely move at EVL cell borders (Supplementary Movie 4).

Furthermore, DCL cells often presented actin brushes at transient contacts with EVL cell borders (Supplementary Fig. 6; Supplementary Movie 5) indicating their ability to sense these regions.

Cells treated with Triton showed significant reduction of phospho-ERM and its colocalization with Dsg3, particularly at the cell borders (Supplementary Figure 3a and b).

In the reverse experiment, the local decrease of EVL tension by overexpression of the N-terminal region of the myosin phosphatase target subunit 1 (N-ter-MYPT1), which reduces the level of phosphorylated myosin19 and disrupts the formation of actin cables along EVL cell borders (Supplementary Fig. 10), led to reduced density of DCL cells under the modified EVL cell (Fig. 8b,c).

Indeed, in epiboly of Fundulopanchax gardneri, a semi-annual killifish species that contains a larger number of DCL cells compared to annual killifish, we observe a transient phase of radial cell intercalation that precedes the formation of a cell monolayer, where DCL cells show a preferential distribution towards EVL cell borders (Supplementary Fig. 13).

Rab11a-deficient enterocytes were still capable to elaborate apical brush borders (Supplementary Fig S3B).

Second, we observed an enrichment of binding sites at previously described domain borders (Supplementary Figures S9 and S10).

arf19 mutant alleles arf19-a (SAIL_92_G09, N872822) and arf19-b (SK-24322, N1007173) were obtained from the Nottingham Arabidopsis Stock Centre and the insertion was confirmed by PCR using forward and reverse primers (obtained using T-DNA primer design tool) in combination with LB3 (for arf19-a) and pSKTAIL-L1 (for arf19-b) border primers (Supplementary Table 1).

The main exception is the AChE staining, where 2 zones of high density of fiber staining (zones 4 6 and 4β 5β; Supplementary Fig. 2) border anteroventraly and posteriorly zones of lower density (zones 1 3 and 3β).