After this incubation, the cell suspension was made up to 1 mL with sterile water. Analysis was performed using an EPICS XL-MCL flow cytometer (Beckman-Coulter, USA) equipped with an argon-ion laser emitting a 488 nm beam at 15 mW. An acquisition protocol was defined after measuring background fluorescence from non-treated BY4741 S. cerevisiae strain, and Δssd1 cells treated with 30 μM FITC-PAF26. Data (20,000 cells/sample) were
analyzed with the DZNeP Expo32 software included in the system acquisition. Acknowledgements S. cerevisiae strain RAY3A and derivatives were kindly provided by Dr. www.selleckchem.com/products/azd5582.html José I. Ibeas (Centro Andaluz de Biologia del Desarrollo, CSIC/Universidad Pablo de Olavide, Sevilla, Spain) to whom we also acknowledge suggestions to the work. We acknowledge the Instituto de Biología Molecular y Celular de Plantas (UPV-CSIC, Valencia, Spain) and M. Dolores Gómez from its microscopy core facility for the use of the confocal microscope. We also acknowledge Drs. José E. Perez-Ortín and José García-Martínez (Laboratory of DNA Chips, Universitat de Valencia, Spain) for advice and suggestions with the macroarray hybridizations and analyses. We appreciate the technical assistance of M. José Pascual (IATA-CSIC), and
the critical review of Adokiye Berepiki (University of Edinburgh, UK). The work was funded by grants BIO2006-09523 and BIO2009-12919 from the Ministry of Science and Innovation (Spain) and ACOMP/2009/080 from Generalitat Valenciana. BLG was hired by the “”Ramón y Cajal”"
program (MEC, Spain), and MG by the JAE-DOC postdoc program (CSIC). Electronic BVD-523 price supplementary material Additional file 1: Sensitivity of S. cerevisiae strains to peptides PAF26 and Melittin. Sensitivity assays of S. cerevisiae strains RAY3A, BWG7a, FY1679, mafosfamide and BY4741 (105 or 104 CFU/mL) to different concentrations of peptides PAF26 and Melittin, at two different assay temperatures. (PDF 444 KB) Additional file 2: Transcriptome analysis of S. cerevisiae FY1679 after exposure to peptides PAF26 and Melittin. Excel File showing the annotation, signal intensity, processing and statistical significance of expression change for each DNA probe in the GPL4565 array. (XLS 4 MB) Additional file 3: Representative S. cerevisiae genes that change expression after exposure to peptides PAF26 and Melittin. Excel File showing lists of genes with the most significant induction/repression that are common or specific after exposure to peptides PAF26 and/or Melittin. (XLS 72 KB) Additional file 4: Non-redundant global GO annotation analyses of S. cerevisiae genes differentially expressed upon peptide treatment. Excel File showing lists of GO annotation terms significantly over- or under-represented among genes induced or repressed after exposure to peptides PAF26 and/or Melittin. (XLS 414 KB) Additional file 5: Sensitivity of gene deletion mutants of S.