FIG. 2.
The peroxisomal membrane protein Pex3p is mislocalized to the cytosol and undefined tubular-vesicular structures in cells repressed for SEC20, SEC39, or DSL1. THC-SEC strains with genomically integrated PEX3-GFP were incubated for 18 h in YEPD (−doxycycline). Repression of TetO7 promoter-regulated genes was achieved by addition of doxycycline to a concentration of 10 μg/ml in YEPD (+doxycycline). Cells were fixed with formaldehyde, images were captured by confocal microscopy, and a maximum-intensity projection was created from a deconvolved 3D data set using Huygens software. Black-and-white images were generated using the “Blend” viewing mode in Imaris (Bitplane) to enhance the total fluorescent signal located within the cell compared to the maximum-intensity projection. Repression of SEC20, SEC39, and DSL1 resulted in redistribution of Pex3p-GFP to the cytosol and undefined tubular-vesicular structures (arrows). Bar, 5 μm.