Quercetin activates the cell wall integrity signaling pathway. Rita Vilaça Vanda Mendes Marta Vaz Mendes Laura Carreto Maria Amélia Amorim Victor de Freitas Pedro Moradas-Ferreira Nuno Mateus Vítor Costa 10.1371/journal.pone.0045494.g005 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> 2012-09-18 01:45:41 activates signaling