Figure 1

Activation of the Pmk1 pathway in response to glucose deprivation. A. Strain MI200 (Pmk1-Ha6H) was grown in YES medium plus 7% glucose to early-log phase and transferred to the same medium with 3% glycerol (upper panel), 2.5% glycerol plus 1% glucose (middle panel) or 2.8% glycerol plus 0.5% glucose (lower panel). Aliquots were harvested at timed intervals and Pmk1 was purified by affinity chromatography. Either activated or total Pmk1 were detected by immunoblotting with anti-phospho-p44/42 or anti-HA antibodies, respectively. B. Strain MI200 was grown in YES medium plus 7% glucose to early-log phase in the presence of 30 mM NAC and resuspended in the same medium with 3% glycerol. Both activated and total Pmk1 were detected as described above.