Figure S4.
Ectopically localized Ent1p, Ent2p, and Sla2p cannot recruit Pan1p. (A) Schematic diagram of the inducible peroxisome redistribution assay. Ent1p, Ent2p, or Sla2p tagged with GFP and FKBP was co-expressed with the peroxisome-targeted Pex-FRB-mCherry fusion protein. Addition of rapamycin causes heterodimerization of FRB and FKBP and thereby recruits Ent1p, Ent2p, or Sla2p to the peroxisome surface. (B) Cells expressing Ent1-, Ent2, or Sla2-GFP-FKBP and Pex-FRB-mCherry were grown to log phase in YPD medium at 25°C, and each image pair was acquired before or at 30 min after rapamycin addition. White arrowheads indicate examples of co-localization. (C) Quantification of the percentages of the indicated GFP-FKBP tagged proteins co-localizing with Pex-FRB-mCherry-labeled peroxisomes before and after rapamcyin addition. Data show the mean ± SD from three experiments (n = 50 puncta for each experiment). (D and G) Schematic diagrams of the inducible peroxisome redistribution assay. Ent1p, Ent2p, or Sla2p tagged with FKBP was co-expressed with Pex-FRB-mCherry in cells expressing (D) GFP-tagged Pan1p or (G) Abp1p. (E and H) Localization of Pex-FRB-mCherry and Pan1-GFP (E) or Abp1-GFP (H) in the presence or absence of rapamycin. Cells were grown to log phase at 25°C and each image pair was acquired before or at 30 min after rapamycin addition. White arrowheads indicate localization of Pex-FRB-mCherry-labeled peroxisome. (F and I) Quantification of the percentages of Pan1-GFP (F) or Abp1-GFP (I) co-localizing with Pex-FRB-mCherry labeled peroxisomes before and after rapamycin addition. Data show the mean ± SD from at least three experiments (n ≥ 50 puncta for each experiment). ***, P value < 0.001, **, P value < 0.01, ns, non-statistically significant, unpaired t test with Welch’s correction. Scale bars, 2.5 μm.

Ectopically localized Ent1p, Ent2p, and Sla2p cannot recruit Pan1p. (A) Schematic diagram of the inducible peroxisome redistribution assay. Ent1p, Ent2p, or Sla2p tagged with GFP and FKBP was co-expressed with the peroxisome-targeted Pex-FRB-mCherry fusion protein. Addition of rapamycin causes heterodimerization of FRB and FKBP and thereby recruits Ent1p, Ent2p, or Sla2p to the peroxisome surface. (B) Cells expressing Ent1-, Ent2, or Sla2-GFP-FKBP and Pex-FRB-mCherry were grown to log phase in YPD medium at 25°C, and each image pair was acquired before or at 30 min after rapamycin addition. White arrowheads indicate examples of co-localization. (C) Quantification of the percentages of the indicated GFP-FKBP tagged proteins co-localizing with Pex-FRB-mCherry-labeled peroxisomes before and after rapamcyin addition. Data show the mean ± SD from three experiments (n = 50 puncta for each experiment). (D and G) Schematic diagrams of the inducible peroxisome redistribution assay. Ent1p, Ent2p, or Sla2p tagged with FKBP was co-expressed with Pex-FRB-mCherry in cells expressing (D) GFP-tagged Pan1p or (G) Abp1p. (E and H) Localization of Pex-FRB-mCherry and Pan1-GFP (E) or Abp1-GFP (H) in the presence or absence of rapamycin. Cells were grown to log phase at 25°C and each image pair was acquired before or at 30 min after rapamycin addition. White arrowheads indicate localization of Pex-FRB-mCherry-labeled peroxisome. (F and I) Quantification of the percentages of Pan1-GFP (F) or Abp1-GFP (I) co-localizing with Pex-FRB-mCherry labeled peroxisomes before and after rapamycin addition. Data show the mean ± SD from at least three experiments (n ≥ 50 puncta for each experiment). ***, P value < 0.001, **, P value < 0.01, ns, non-statistically significant, unpaired t test with Welch’s correction. Scale bars, 2.5 μm.

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