MYO18B promotes focal adhesion maturation. (A) Left: Representative results of wound healing in WT and MYO18B-KO HeLa cells. DIC images were acquired immediately and 24 h after cells were scratched. Bars, 200 µm. Right: Quantitative result of wound healing assay in WT and MYO18B-KO HeLa cells. Data was acquired at 0, 4, 8, 12, and 24 h after scratching (n = 3 images, mean ± SD, Two-way ANOVA, **P ≤ 0.01). Related to Fig. 4. (B) Left: Representative images of WT and MYO18B-KO U2OS cells stained with vinculin. Bars, 10 µm. Right: Quantitative result of vinculin puncta >0.5 µm² in WT and MYO18B-KO U2OS cells (mean ± SD; n = 15 images; N = 3 biological replicates; unpaired two-tailed t test; ***P ≤ 0.001). Related to Fig. 4. (C) Representative CLSM images of WT and MYO18B-KO HeLa cells stained with vinculin. Bar, 10 µm. Related to Fig. 4. (D) Raw fluorescent intensity detected in HEX activity assessment of MYO18B-KO HeLa cells expressing different EGFP-tagged MYO18B truncates. The fluorescent intensity of intracellular fraction was normalized with protein amount (mean ± SD; n = 3, independently analyzed samples; One-way ANOVA with Dunnett correction; ns, P > 0.05; *P ≤ 0.05; **P ≤ 0.01; ***P ≤ 0.001; ****P ≤ 0.0001). Related to Fig. 5. (E) Representative CLSM images of MYO18B-KO HeLa cells expressing different EGFP tagged MYO18B truncates stained with vinculin. Bars, 10 µm. Related to Fig. 5.