Fig. 6: Dual targeting of Pxp2 to mitochondria and peroxisomes.
From: Peroxisomal targeting of a protein phosphatase type 2C via mitochondrial transit
a Subcellular localization of Pxp2-RFP (red) and Pxp2-RFP-PTS (red) was determined using fluorescence microscopy. Ant1-YFP (green) and Tim50-CFP (blue) were used to label peroxisomes and mitochondria, respectively. White color represents overlap of all three signals. Scale bar represents 5 µm. b Correlation of Pxp2-RFP or Pxp2-RFP-PTS with Ant1-YFP was analyzed. PCC refers to Pearson’s correlation coefficient. Quantifications are based three independent experiments (n = 3). Each experiment is represented by a different color. Error bars represent standard error of the mean. P-values were calculated with a two-tailed unpaired Student’s t-test. c Subcellular localization of Pxp2-RFP and Pxp2-RFP-PTS was determined using density gradient centrifugation. Twelve fractions, collected from the top of the gradient, were analyzed by SDS–PAGE and western blot. d Twenty five amino acids from the N-terminus of Pxp2 were fused to RFP. Pxp21–25-RFP (magenta) was expressed in a strain also containing Tim50-YFP (cyan). The resulting strain was analyzed by fluorescence microscopy. White color indicates colocalization. Scale bar represents 5 µm. e Quantification of the fraction of peroxisomes contacting mitochondria (PxM) in relation to the total peroxisome count (PxT) of cells co-expressing Pxp2-RFP or Pxp2-RFP-PTS with Ant1-YFP, and the mitochondrial membrane protein Tim50 fused to CFP. Association of Ant1-YFP containing foci with Tim50-CFP was counted. Quantifications are based on three independent experiments (n = 3). Each experiment is represented by a different color. Error bars represent standard error of the mean. P-values were calculated with a two-tailed unpaired Student’s t-test. Source data are provided in the Source data file.
