Fig. 5

Analysis of genetically modified yeast cells. (a) Yeast IB1- and IB2-derivative strains, carrying one chromosomal copy of the YSD-yCup1 transgene replacing ADE2 gene, were analyzed by immunofluorescence microscopy to visualize the localization of the fusion protein. For each strain, micrographs are reported of the differential interference contrast (DIC), the immunofluorescence signal (AF555), the nuclear DNA staining (DAPI), and the merged image. Images are representative of three biological replicates for each yeast strain. Scale bar: 5 μm. (b) 5 × 10⁸ yeast cells of the IB1 and IB2 strains, either unmodified (WT), or carrying one chromosomal copy of the yCup1 chimeric transgene (ade2D::YSD-yCup1), were checked for Cu²⁺ binding by ICP-EOS and quantification data are reported in the graphs (as nmoles of Cu²⁺ / mg CDW). The ratio between values of modified and parental strains (YSD/WT) is indicated for each strain. Each column represents the mean ± SEM of 5 independent experiments (n = 5; *p < 0,05, **p < 0,01, Mann Whitney T-test was applied for statistical analysis)