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. 1999 May;10(5):1395–1407. doi: 10.1091/mbc.10.5.1395

Figure 2.

Figure 2

Nuclear accumulation of Spc1 requires Pim1 function. (A) GD1942 (spc1–12myc) cells were exposed to osmotic stress (0.6 M KCl in YES medium). Immunolocalization with antibodies to myc revealed that Spc1 became concentrated in the nucleus at the 10-min time point. Nuclei were visualized with the DNA stain DAPI. (B) A pim1-d1 spc1–12myc strain was incubated at the restrictive temperature of 36°C for 3 h and stressed with 0.6 M KCl. Spc1 failed to accumulate in the nucleus in response to stress. (C) Phosphorylation of Spc1 on tyrosine-173 was detected by immunoblot analysis using antibodies to the phosphorylated tyrosine-182 epitope of human p38 (α-p38P). Osmotic stress caused tyrosine phosphorylation of Spc1 in wild-type (WT) and pim1-d1 cells. Total Spc1–12myc protein was detected with antibodies to myc. (D) Exponentially growing wild-type cells (PR109) and pim1-d1 mutant were stressed with KCl and harvested at the indicated time, and total RNA was extracted for Northern analysis of pyp2+ or leu1+ mRNA (loading control).