Figure 2.
Pan1p recruits other NPFs and triggers actin polymerization. (A) Localization of Abp1-GFP and Pex-FRB-mCherry before or at 1 min 45 s after CK666 treatment in the presence of rapamycin (left and center image pairs). Right image pair in A was acquired at 1 min 35 s after washing away CK666. (B) Data show the mean ± SD from three experiments (n = 50 puncta for each experiment). Different letters indicate significant difference between indicated times at P < 0.001, one-way ANOVA with Turkey’s post-hoc test. (C) Schematic diagrams of the inducible peroxisome redistribution assay. Pan1-GFP-FKBP was co-expressed with Pex-FRB-mCherry in cells expressing Abp1-CFP. (D) Localization of Pan1-GFP-FKBP, Pex-FRB-mCherry, and Abp1-CFP at 30 min after rapamycin addition. (E) The time series of each protein in the boxed area (D) are shown in the lower panels. Blue or red arrowheads indicate the timing of the appearance or disappearance respectively of Abp1-CFP signal. Quantification of fluorescence intensities for each protein in the boxed area is shown in the right panel. (F) Averaged fluorescence intensities as a function of time for each protein (n = 10 independent peroxisomes). (G) FKBP-tagged Pan1p was co-expressed with Pex-FRB-mCherry in cell expressing indicated GFP-tagged proteins. (H) Each image pair was acquired at 30 min after rapamycin addition. White arrowheads indicate examples of co-localization. (I) 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). ***, P value < 0.001, **, P value < 0.01, unpaired t test with Welch’s correction. Scale bars, 2.5 μm.

Pan1p recruits other NPFs and triggers actin polymerization. (A) Localization of Abp1-GFP and Pex-FRB-mCherry before or at 1 min 45 s after CK666 treatment in the presence of rapamycin (left and center image pairs). Right image pair in A was acquired at 1 min 35 s after washing away CK666. (B) Data show the mean ± SD from three experiments (n = 50 puncta for each experiment). Different letters indicate significant difference between indicated times at P < 0.001, one-way ANOVA with Turkey’s post-hoc test. (C) Schematic diagrams of the inducible peroxisome redistribution assay. Pan1-GFP-FKBP was co-expressed with Pex-FRB-mCherry in cells expressing Abp1-CFP. (D) Localization of Pan1-GFP-FKBP, Pex-FRB-mCherry, and Abp1-CFP at 30 min after rapamycin addition. (E) The time series of each protein in the boxed area (D) are shown in the lower panels. Blue or red arrowheads indicate the timing of the appearance or disappearance respectively of Abp1-CFP signal. Quantification of fluorescence intensities for each protein in the boxed area is shown in the right panel. (F) Averaged fluorescence intensities as a function of time for each protein (n = 10 independent peroxisomes). (G) FKBP-tagged Pan1p was co-expressed with Pex-FRB-mCherry in cell expressing indicated GFP-tagged proteins. (H) Each image pair was acquired at 30 min after rapamycin addition. White arrowheads indicate examples of co-localization. (I) 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). ***, P value < 0.001, **, P value < 0.01, unpaired t test with Welch’s correction. Scale bars, 2.5 μm.

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