Failure of BiFC to segregate with peroxisomes in mating cells, haploid cells, and diploid cells. (A) WT MatA Pex14-VC cells containing red-PTS1 on a plasmid were mated with WT Matα Pex13-VN cells and imaged 24 h after mating. Two examples are shown. (B) BiFC between Pex2-VN and Pex14-VC in WT haploid cells expressing HcRed-PTS1 from a plasmid. (C and D) VPS1 was disrupted in haploid cells double tagged as indicated, and peroxisomes were visualized using Ant1-mCherry. Two examples of each are shown. (E and F) Pex13-VN/14-VC BiFC (E) versus Pex13-GFP and Pex14-GFP (F) in diploid vps1 cells expressing HcRed-PTS1 from a plasmid. (G) Video stills from Video 2 of Pex13-VN/Pex14-VC diploid vps1 cells expressing HcRed-PTS1 from a plasmid. VN, VC, and GFP tags were genomically integrated (i.e., a WT copy in addition to tagged protein was present in diploid cells). Red arrowheads indicate peroxisomes or parts of peroxisomes without a BiFC signal. White arrowheads indicate punctate BiFC signals colocalizing with the elongated peroxisomes. White arrowheads in G indicate peroxisome with BiFC signal from which a peroxisome without BiFC signal splits off (red arrowheads). Bars, 5 µm.