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. 1997 Jun 30;137(7):1495–1509. doi: 10.1083/jcb.137.7.1495

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Immunofluorescence localization of Sec4p in sec mutants. Cells were grown overnight in YP dextrose medium at 25°C and then shifted to 37°C for 1 h, fixed, and labeled with αSec4 antibody. The effect of cycloheximide (cyc) and azide treatment on wild-type cells was tested by adding either agent to the culture immediately before shift to 37°C. sec2-78, NY1529; myo2-66, NY1005; act1-3, NY278; sec1-1, NY8; sec3-2, NY45; sec4-8, NY28; sec5-24, NY30; sec6-4, NY18; sec8-9, NY43; sec9-4, NY32; sec10-2, NY36; sec15-1, NY39; wild-type, NY10; sec12-4, NY738; sec16-2, NY416; bos1-1, NY1282 (see Table I for strain genotypes). Wild-type cells and most late-acting sec mutants show immunolabeling preferentially in the bud. In sec2-78, myo2-66, and act1-3, diffuse staining for Sec4p is observed. Also, in sec4-8, ER to Golgi sec mutants (sec12-4, sec16-2, bos1-1), and after treatment of wild-type cells with cycloheximide or azide, no concentration of immunolabeling in the bud is found.