Monitoring of Rst2 transcriptional activity in living cells by using the <i>Renilla</i> luciferase reporter assay.

<p>(A) Glucose deprivation induced a marked increase in transcriptional activation. Wild-type cells harboring the reporter plasmid were cultured and assayed as described under “Materials and Methods”. GR (light gray line) indicates that the cells were resuspended in glucose-rich medium (GR). GD (dark gray line) indicates that the cells were resuspended in low glucose medium to induce glucose deprivation (GD). Y-axis values are the ratio of relative light units (RLU) of each sample to that of wild-type cells in GR at 150 minutes. The data shown are representative of multiple experiments. (B) Glucose deprivation-induced transcriptional activation is completely abolished in Δ<i>rst2</i> cells. The Δ<i>rst2</i> cells harboring the reporter plasmid were cultured and assayed as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0078012#pone-0078012-g001" target="_blank">Figure 1A</a>. (C) Rst2 is specifically activated by glucose deprivation. Wild-type cells harboring the reporter plasmid were treated with GR, GD, 1 mM H₂O₂, 300 mM KCl or 1 mM CdCl₂ as indicated. Area under the curve (AUC) is expressed as a percentage of RLU of wild-type cells in GD from 0 to 300 minutes. Error bars, mean ± S.D. (n ≥ 3).