Figure 2

Effect of LI0758 on yeast growth. A. Examples of growth defects in yeast strains expressing LI0758 proteins. The yeast strains BY4741 and W303-1A, each transformed with an empty vector, RipI (serving as a positive control), and LI0758 under the regulation of the GAL1 promoter, were cultivated to late logarithmic phase in medium containing 2% dextrose. Subsequently, tenfold serial dilutions (ranging from 10⁰ to 10⁴) were spotted onto agar plates containing either glucose (as a repressor) or galactose (as an inducer), supplemented with 0.5 M NaCl or 0.8 M sorbitol. The plates were then incubated at temperatures of 30 °C or 37 °C. The experiments were conducted in triplicate, with consistent results obtained. B. Liquid growth assay and western blot of yeast strains BY4741. Three clones transformed with LI0758 (denoted as clones 1, 2, and 3) were randomly selected. These clones were first subjected to expression level analysis via western blotting. Subsequently, their growth over time was analyzed in a liquid growth assay. The error bars represent the standard deviation of the mean values from three independent experiments. C. Liquid growth assay and western blot of yeast strains W303-1A. Three clones transformed with LI0758 (denoted as clones 1, 2, and 3) were randomly selected. These clones were first subjected to expression level analysis via western blotting. Subsequently, their growth over time was analyzed in a liquid growth assay. The error bars represent the standard deviation of the mean values from three independent experiments.