FIG 9.

Cells lacking Reg2 exhibit delayed recovery from prolonged glucose deprivation. (A) Wild-type and reg2Δ cells were grown to mid-log phase in YEP containing 2% glucose and then shifted to YEP without glucose for 24 h. After this starvation period, the cultures were shifted (at an initial OD₆₀₀ of 0.15) to YEP containing 2% glucose, and the OD₆₀₀ of the cultures was monitored. The values are averages for three cultures of each genotype. The error bars indicate standard errors. (B) Wild-type (REG2, +) and reg2Δ (REG2, Δ) cells were grown to mid-log phase in YEP medium containing 2% glucose and shifted to YEP containing no glucose for 24 h, and glucose was then added at 2% (+Glu) for either 15 min (+15′), 20 min (+20′), or 25 min (+25′). The levels of phosphorylated Thr210 (P-T210) and total Snf1 protein (Total) were analyzed by immunoblotting. (C) Wild-type and reg2Δ cells were grown to mid-log phase in YEP medium containing 2% glucose, mixed in equal proportions, and subjected to 10 cycles consisting of a 24-h glucose starvation phase and a 5- to 6-h glucose replenishment phase, and the proportion of reg2Δ cells was determined as described in Materials and Methods. In all the experiments, the strains were MKY323 (WT) and MMY9 (reg2Δ).