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. 1997 Aug 11;138(3):517–529. doi: 10.1083/jcb.138.3.517

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Vacuolar inheritance analysis of vam3^tsf cells. TDY1 (vam3Δ) cells carrying either complementing VAM3 plasmid (pVAM3.414) or vam3^tsf plasmid (pVAM3-6.414) were grown to exponential phase and then labeled with 32 μM FM4-64 for a period of 45 min. Cultures were chased with excess YPD for 1 h at 26°C and then split into equal aliquots. The aliquots were chased for an additional 2 h at either 26°C or 38°C. The cultures were then examined by fluorescence and Nomarski microscopy for the presence of vacuolar segregation structures. (White arrowheads) Segregation structures.