1. Lead induces oxidative stress and phenotypic markers of apoptosis in Saccharomyces cerevisiae (Applied Microbiology and Biotechnology) Jurrian Vanden Bussche and Eduardo V. Soares 1 1 Author for correspondence: Eduardo V. Soares Bioengineering Laboratory, Chemical Engineering Department, Superior Institute of Engineering from Porto Polytechnic Institute, Rua Dr António Bernardino de Almeida, 431, 4200-072 Porto, Portugal e-mail: evs@isep.ipp.ptevs@isep.ipp.pt Tel: 351-22-8340500 Fax: 351-22-8321159 1

2. Fig. S1. Membrane disruption in cells of S. cerevisiae NCYC 1214 heated at 65 ºC, during one hour (control cells). Cells were stained with Oxonol or propidium iodide (PI), as described in material and methods. Oxonol can freely enter in the cells with depolarized membrane (a). Cells with compromised membrane incorporate PI (c). Fluorescence micrographs (a and c); phase contrast micrographs of the same cells (b and d). a c b d 2

3. Fig. S2. Membrane disruption in cells of S. cerevisiae NCYC 1214 heated at 65 ºC, during one hour (control cells). A double staining protocol with Oxonol and PI was carried out, as described in material and methods. Fluorescent micrographs of the cells observed with filter set GFP (a) or I3 (b); phase contrast micrograph of the same cells (c). a b c 3