Three NPFs colocalize with Arp2/3 at different cellular and subcellular locations. (A) Covisualization of Arp2/3 with WAVE, WASP, and WASH in gastrulation stage embryos using endogenously tagged alleles. White arrowheads point to Ea and Ep cells. Scale bar: 5 µm. (B) Quantification of colocalization between Arp2/3 and NPFs, with box plots reporting Pearson correlation coefficients at Ep-P₄ contacts, other cell–cell contacts, and the cytoplasm. Statistical tests for experiments in B were chosen based on the normality and variance of the data: For WAVE, one-way ANOVA was followed by Post-hoc Tukey’s tests; for WASP and WASH, Kruskal–Wallis test was followed by post-hoc Dunn’s test (center line, median; box, interquartile range (IQR); whiskers, min/max range; n ≥ 10 embryos; *P < 0.05, **P < 0.01, ****P < 0.0001.