A block in pexophagy restores BiFC signals in pex1 and pex6 cells. (A) BiFC between Pex2-VN and Pex14-VC (genomically tagged) in haploid cells. Cells were transformed with a plasmid expressing Ant1-mCherry and grown on a glucose-containing medium to allow BiFC signals to develop. Mean number of Ant1-mCherry puncta per cell + SD were as follows: WT, 2.5 ± 1.6; atg36, 3.5 ± 2.3; pex1, 0.55 ± 0.6; pex1/atg36, 1.4 ± 0.7; pex6, 0.6 ± 0.6; and pex6/atg36, 1.6 ± 0.9. Percentage of cells showing colocalization between BiFC and Ant1 were as follows: WT, 84%; atg36, 89%; pex1, 95%; pex1/atg36, 93%; pex6, 98%; and pex6/atg36, 90%. At least 100 cells were analyzed. (B) Western blot showing Pex14-VC in strains as indicated. PGK1 was used as a loading control. (C) BiFC between mitochondrial and peroxisomal proteins in WT and pex1 cells as indicated. A singe slice of each sample was captured so that the fluorescence intensity in the images reflects the BiFC signal strength in the samples. Bars, 5 µm.