USP45 depletion accelerates recovery of acidification and actin patch structure formation after removal of actin polymerization inhibitor, LatA. (A) Immunofluorescent analysis of LysoTracker (LTK) and actin structure (phalloidin) in control and USP45 knockdown HeLa cells. Cells were treated with LatA (200 nM, 2 h) followed by replacement of fresh medium for 15 min. Scale bar, 20 μm (original) and 5 μm (zoom-in). (B) Quantification of LTK dot number and LTK-phalloidin colocalization in A. Data are shown as mean ± SEM in n = 3, ≥30 cells. Significance was determined using one-way ANOVA and Tukey’s multiple comparisons test; ****P < 0.0001.