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>