RNAi against each NPF leads to Arp2/3 reduction at different cellular and subcellular locations. (A) Micrographs from time-lapse movies depicting localization of Arp2/3 in control and WAVE, WASP, or WASH RNAi-treated embryos from a lateral view. White arrowheads point to Ea and Ep cells. The diagrams underneath each micrograph highlight the observed Arp2/3 localization in E and neighboring cells. (B) Violin plots reporting changes in Arp2/3 localization at Ep-P₄ contacts, other cell-cell contacts, and the cytoplasm upon RNAi depletion of WAVE, WASP, and WASH (center dot, mean; vertical line, s.d.; outline, the distribution of the data; n ≥ 10 embryos). (C) Violin plots reporting changes in the differences between the maximum and minimum gray values of the Arp2/3 signal along the line scan upon RNAi depletion of WAVE, WASP, and WASH (center dot, mean; vertical line, s.d.; outline, the distribution of the data; n ≥ 10 embryos). All measurements were collected 6 min after the division of neighboring mesoderm precursor cells (MSx). Statistical tests for experiments in B and C were chosen based on the normality and variance of the data. (B) For WAVE (other cell contacts [control versus RNAi]), unpaired t test with Welch’s correction; for WAVE (others), unpaired t test; for WASP (Ep–P4 contacts [control versus RNAi]), unpaired t test with Welch’s correction; for WASP (others), unpaired t test; for WASH (Ep–P4 contacts [control versus RNAi]), unpaired t test; for WASH (other cell contacts [control versus RNAi]), unpaired t test with Welch’s correction; for WASH (cytoplasm), Mann-Whitney test. (C) For WAVE and WASP, unpaired t test; for WASH, Mann–Whitney test. *P < 0.05, **P < 0.01, ****P < 0.0001. Scale bar: 5 µm.