Inp1p interacts with cytosolic domain of Pex3p in vivo. (A–C) Split-GFP analysis between Inp1p and Pex3p in WT cells. Tagged proteins were expressed under control of the GAL1 promoter for 4 h (short) or 8 h (long) and scored for the presence and intensity of fluorescence (A). −, no signal; +, faint; +++, strong. (B) Selected images of WT cells induced for 4 or 8 h. (C) WT cells were induced to express Inp1p-GFP-N and Pex3p-GFP-C for 4 h, followed by mating with pex3Δ cells expressing HcRed-PTS1, and imaging 2 h after mating. GFP and HcRed signals overlap in mated cell (arrow). (D) The expression of a chimeric protein consisting of the cytosolic domain of Pex3p fused at its N terminus to Tom70p and tagged at its C terminus with mRFP (mito-Pex3p-mRFP) was induced on galactose for 3 h in pex3Δ cells expressing Inp1p-GFP under control of its endogenous promoter. Image shows two budding cells, one of which is expressing mito-Pex3p-RFP that recruits Inp1p-GFP. The strains were grown on selective medium and examined by epifluorescence and phase contrast. Multiple epifluorescence images were acquired in the z axis and flattened into a single image. The brightfield image is blue in the merged pictures. Bar, 5 µm.