10.1371/journal.pone.0045494.g005 Rita Vilaça Rita Vilaça Vanda Mendes Vanda Mendes Marta Vaz Mendes Marta Vaz Mendes Laura Carreto Laura Carreto Maria Amélia Amorim Maria Amélia Amorim Victor de Freitas Victor de Freitas Pedro Moradas-Ferreira Pedro Moradas-Ferreira Nuno Mateus Nuno Mateus Vítor Costa Vítor Costa Quercetin activates the cell wall integrity signaling pathway. Public Library of Science 2012 activates signaling 2012-09-18 01:45:41 Figure https://plos.figshare.com/articles/figure/_Quercetin_activates_the_cell_wall_integrity_signaling_pathway_/246341 <p>(A) Phospho-Slt2 protein levels. <i>S. cerevisiae</i> BY4741 cells were grown to exponential phase and treated with 300 µM of quercetin or equal volume of DMSO for the indicated time periods. Proteins were isolated, separated by SDS-PAGE, blotted into a membrane and incubated with anti-phospho-p44/42 antibody that detects dually phosphorylated Slt2p or with anti-actin antibody (loading control), as described in <i><a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0045494#s4" target="_blank">Materials and Methods</a></i>. A representative blot is shown. (B) Rlm1p-transcription factor activity. Exponentially growing <i>S. cerevisiae</i> BY4741 cells transformed with a <i>pRLM1-LacZ</i> reporter construct were treated with 300 µM of quercetin or equal volume of DMSO (control) at the indicated times. β-galactosidase activity was determined as described in <i><a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0045494#s4" target="_blank">Materials and Methods</a></i> and expressed as percentage of control cells. (C) Resistance to zymolyase digestion. <i>S. cerevisiae</i> BY4741 cells were treated with 300 µM of quercetin or equal volume of DMSO for 15 min and incubated with 0.25 U/ml zymolyase at 37°C. Cell lysis was determined spectrophotometrically at 600 nm over time. (D) Hydrogen peroxide resistance of <i>slt2</i>Δ cells was assessed as described in legend to <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0045494#pone-0045494-g004" target="_blank">Figure 4D</a>. Values are mean ± SD of at least three independent assays. **p<0.01; *p<0.05 (unpaired t-test).</p>