Atg18-mN G localizes predominantly to the daughter vacuole periphery. (A–D) Time-lapse microscopy–based analysis of Atg18-mNG vacuole periphery localization during the cell cycle in cells containing Vph1-3xmCherry (MUK089). Selected time points from a representative time-lapse series are shown (A) and mean PCC values of Atg18-mNG colocalization with Vph1-3xmCherry are plotted (B). Data are aligned with respect to the timing of vacuole segregation (t = 0) in B and the error bars show the standard deviation. White arrowhead in A indicates the end of vacuole segregation (t = 0). Timing of Atg18-mNG appearance on the daughter vacuole with respect to completion of vacuole segregation (C) and persistence of Atg18-mNG signal on both vacuoles (D) are plotted. Ordinary one-way ANOVA was performed with Fisher’s LSD test. ****P < 0.0001. Data come from three independent time-lapse experiments. (E) Atg18-mNG daughter vacuole localization with respect to the timing of anaphase onset and spindle breakdown in mCherry-Tub1–containing cells (MUK116). Time-lapse series of a representative cell are shown. Yellow arrowheads mark the start of anaphase onset and spindle breakdown, respectively. The graph shows Atg18-mNG signal appearance (arrival) and disappearance from the daughter vacuole (departure) relative to the timing of anaphase onset and spindle breakdown, respectively. Data come from two independent time-lapse experiments. Scale bars are 3 μm, n: sample size. DV: daughter vacuole, MV: mother vacuole. Circles in scatter plots are values of individual cells. Black lines in scatter plots represent the mean and standard deviation. m: mother, d: daughter.