Direct anchoring of Las17-, Myo3-, or Myo5-FKBP to the peroxisome does not recruit Pan1p to the peroxisome. (A) Cells expressing Pan1-GFP, Pex-FRB-mCherry, and FKBP-tagged Las17p (left panels), Myo3p (center panels), or Myo5p (right panels) were grown to log phase at 25°C, and each image pair was acquired before or 30 min after rapamycin addition. (B) The percentage of Pan1-GFP co-localizing with Pex-FRB-mCherry-labeled peroxisomes in the presence of the indicated other FKBP-fusion protein before and after rapamycin addition. Data show the mean ± SD from at least three experiments (n > 30 puncta for each experiment). (C) FKBP-tagged Las17p, Myo3p, or Myo5p was co-expressed with Pex-FRB-mCherry and Abp1-GFP. (D) Each image pair was acquired at 30 min after rapamycin addition. (E) Quantification of the percentage of each GFP-tagged protein localizing at Pex-FRB-mCherry-labeled peroxisomes. Data show the mean ± SD from three experiments (n = 50 puncta for each experiment). ns, non-statistically significant, unpaired t test with Welch’s correction. Scale bars, 2.5 μm.