Dynamic localization of actin disassembly factors at Pan1p-anchoring peroxisomes. (A) Spatial-temporal recruitment of Prk1p, Cof1p, and Srv2p. Timing of appearance and disappearance of actin disassembly factors are shown below. (B) Localization of Prk1-GFP, Pex-FRB-mCherry, and Abp1-mTur2 before or at 30 min after rapamycin addition. (C) Quantification of the percentage of Prk1-GFP localizing at Pex-FRB-mCherry-labeled peroxisomes. Data show the mean ± SD from three experiments (n = 50 puncta for each experiment). (D) The time series of each protein in the boxed area in B are shown. Blue or red arrowheads indicate the timing of the appearance or disappearance of Abp1-CFP signal. (E and H) The localization of GFP-Cof1 (E) or Srv2-GFP (H), Abp1-mTq2, and Pex-FRB-mCherry at 30 min after rapamycin addition. A time series of a single peroxisome in the boxed area is shown in the lower panels. Blue or red arrowheads indicate the timing of the appearance or disappearance Abp1-CFP signal. (F and I) Averaged fluorescence intensities as a function of time for each protein (n = 10 independent peroxisomes). (G and J) Quantification of the percentages of GFP-Cof1 (G) or Srv2-GFP (J) localizing at Pex-FRB-mCherry-labeled peroxisomes before and after rapamycin addition. Throughout figure blue or red arrowheads indicate the timing of the appearance or disappearance of the indicated fusion protein’s signal. Data show the mean ± SD from three experiments (n = 100 puncta for each experiment). **, P value < 0.01, ***, P value < 0.01, unpaired t test with Welch’s correction. Scale bars, 2.5 μm.