Functional differences between MYO18B and MYO18A. (A) Representative flow cytometry results of cell-surface LAMP1 level in WT, MYO18Aα-KO HeLa, and MYO18B-KO U2OS cells. Related to Fig. 5. (B) Representative result of HEX activity assay in WT, MYO18Aα-KO HeLa, and MYO18B-KO U2OS cells (n = 3, independently analyzed samples; mean ± SD; One-way ANOVA with Dunnett correction; **P ≤ 0.01; ***P ≤ 0.001; ****P ≤ 0.0001). Related to Fig. 5. (C) Left: Representative CLSM images of WT, MYO18Aα-KO, and MYO18B-KO U2OS cells stained with vinculin. Bars, 10 µm. Right: Quantitative results of vinculin puncta >0.5 µm² in WT MYO18Aα-KO and MYO18B-KO U2OS cells (mean ± SD; n = 15 images; N = 3 biological replicates; unpaired two-tailed t test; **P ≤ 0.01, ****P ≤ 0.0001). Related to Fig. 5. (D) Representative flow cytometry results of cell-surface LAMP1 level in WT, MYO18Aα-KO HeLa and MYO18B-KO U2OS cells treated by Pistop2. Related to Fig. 5. (E) RT-qPCR assessment of MYO18A mRNA level in MYO18B-KO U2OS cells. Expression of MYO18A was normalized with HPRT (n = 3 technical replicates; One-way ANOVA with Dunnett correction; ns, P > 0.05). (F) Representative result of plasma membrane repairing assay in MYO18B-depleted cells expressing EGFP-ACTN1. Related to Fig. 5.