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. 2003 Jun;2(3):510–520. doi: 10.1128/EC.2.3.510-520.2003

FIG. 6.

FIG. 6.

scw1Δ does not rescues SIN mutants by restoring SIN localization and activity. (A and B) The indicated strains were grown in YE at 25°C to log phase and then shifted to 33.5°C for 4 h before being fixed and stained with DAPI and photographed for GFP fluorescence. WT, wild type. (C) The scw1Δ mutation does not promote Sid2p kinase activity. Various strains were harvested either in log phase at 25°C or after being shifted for 4 h to 36°C. The presence (+) or absence (−) of either Sid2-13Myc or the scw1Δ mutation is indicated, as is the presence of SIN mutations. Immune complexes were prepared from lysates with anti-Myc antibody, and then a kinase assay was performed with MBP as an artificial substrate, as previously described (47). Each sample was split in two, and phosphorylation of MBP (upper panels) or Sid2p-13Myc levels (lower panel) were detected by phosphorimager and Western analysis, respectively. IP, immunoprecipitation.