Figure 6.—

MAK1 activity is increased in cot-1(ts). (A) Total soluble protein (100 μg/lane) was extracted from the indicated strains grown in the presence or absence of stress inducers (1 m NaCl, 7 mm H₂O₂). The blot was probed with anti-phospho-ERK (α-P-ERK) and anti-phospho-p38 (α-P-p38) antibodies to detect activated MAK1, MAK2, and OS2 kinase. (B) For the temperature-shift experiments, total soluble protein (50 μg/lane) of the indicated strains grown at 25° and shifted to 37° for 12 hr was extracted and the blot was probed with anti-phospho-ERK (α-P-ERK) antibody (top). To confirm equal loading, the blot was stripped and reprobed with α-tubulin antibody (bottom). lrg-1 is an unrelated temperature-sensitive hyperbranching mutant used as a control.