Exocytic carrier sequestration by EHBP-1 depends on RAB-10 functionality. (A) Domain architecture and subcellular localization of EHBP-1 and Mito-EHBP-1^6AALAA. (B and B′) Confocal images showing colocalization between VIT-2-mNeonGreen-3xFlag^KI and mitochondria. Statistical analysis of colocalization was calculated as Pearson’s correlation coefficient and is shown as the mean ± SD (n = 18 cells from six animals of each genotype; one-way ANOVA test with Dunn’s multiple comparison). (C) TEM images showing the spatial relationship between the yolk and the mitochondria. Yellow and blue indicate the yolk and the mitochondria, respectively. (D and D′) Confocal images showing colocalization between GFP-PLIN2-labeled lipid droplets and lipoproteins in HepG2 cells. Statistical analysis was calculated as Pearson’s correlation coefficient and is shown as the mean ± SD (n = 18 cells; one-way ANOVA test with Dunn’s multiple comparison). (E) Cartoon image showing the liposome tethering assay. (F and F′) The images acquired using a TIRF microscope showing liposomes equipped with RAB-10(GMP-PNP) or RAB-10(GDP) captured by slides immobilized with StrepII-EHBP-1(ΔC2) or StrepII-EHBP-1(ΔC2 ΔCC). Statistical data are shown as box-and-whisker plots with the 10th–90th percentile (n = 24 areas; dots, outliers; boundaries, quartiles; two-way ANOVA with Bonferroni post-test).