FIG. 5.
The heat stress-induced increase in Slm1 and Slm2 phosphorylation is independent of Tor2p. Overnight cultures of wild-type (wt; JK9-3da) and tor2ts (SH121) cells expressing either GST-Slm2 or GST-Slm1 were diluted to an OD600 of 0.2 and grown at 25°C for 4 h. To one set of cultures, FK506 (final concentration, 2 μg/ml) or solvent (90% ethanol/10% Tween 20) was added and cells were shifted to 38°C. After 30 min, galactose (2%) was added to induce protein expression and cells were grown for another 4 h at 38°C. GST fusion proteins were purified from cell extracts and subjected to SDS-PAGE and Western blotting with anti-phospho and anti-GST antibodies.