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

Figure 3.

Figure 3

Immunofluorescence localization of Sec4p in different sec2 alleles and a sec2-78,6-4 double mutant after a short shift to the restrictive temperature. Cells were grown overnight in YP dextrose medium at 25°C and then shifted to 37°C for 10 min, fixed, and labeled with αSec4 antibody. sec12-4, NY738; sec2-41, NY132; sec2-59, NY27; sec2-78, NY1529; wild-type, NY10; sec6-4, NY779; sec2-78,6-4, NY1530. In sec2-41, sec2-59, and sec2-78, Sec4p, immunostaining was concentrated in the bud at 25°C but was diffuse already after 10-min shift to the restrictive temperature. In the ER-to-Golgi mutant sec12-4, diffuse immunolabeling was also already observed after 10 min at 37°C. Both in wild-type and sec6-4, Sec4p immunolabeling was concentrated at the bud both at 25°C and after 10 min at 37°C. The double mutant was constructed as described in Materials and Methods. sec2-78, sec2-78,6-4 shows labeling in the buds at the permissive temperature that is no longer detected after 10 min at the restrictive temperature.