Figure 1. Stress induced Rom2 phosphorylation is Mpk1 dependent.

Exponentially growing cells (Y974) were shifted from 23°C to 38°C (A. or treated with rapamycin at 23°C (B. for indicated duration. Cells were collected and lysed. The expressed Rom2-myc₁₃ and the levels of phospho-Mpk1 in the lysates were detected by western blotting. C. Wild cells expressing Rom2-myc₁₃ (Y974) were grown to early log phase at 23°C and shifted to 38°C. Aliquots of cells were removed at the indicated time points after the shift and lysed. Rom2-myc₁₃ was immunopurified with anti-Myc antibody and treated for 1 hr with alkaline phosphatase (Pase+) in the presence (PI+) or absence (PI−) of 20 mM of β-glycerophosphate. D. Exponentially growing wild type (Y974) and its isogenic mpk1Δ cells expressing Rom2-myc₁₃ (Y973) were shifted from 23°C to 38°C. Aliquots of cells were removed at the indicated time points and lysed. The expressed Rom2-myc₁₃ in the lysates was detected by western blotting.