Figure 7

In vivo activity and function analysis of LI0758. A. Mating efficiency of yeast strains expressing LI0758. Quantitative mating assays were conducted in yeast as described in Materials and methods section. Each assay was conducted in triplicate, with the wild-type strain serving as the reference (set to 100%). The mating efficiency of mutant strains is expressed as a percentage relative to the wild-type. The error bars indicate the standard deviation from the mean values. B. Expression confirmation of ScRce1 and LI0758 by western blot. Western blot analysis was conducted using an anti-Flag antibody to verify the expression of ScRce1 and LI0758. Numbers 1, 2, and 3 indicate three randomly selected clones from yeast transformed with LI0758, which were later used in the mating experiments shown in A. C. Localization of GFP-Ras2 in rce1Δ mutant strains. To compare the localization patterns and intensities of the GFP-Ras2 fusion protein, rce1Δ mutant strains were created that expressed GFP-Ras2 along with either an mCherry empty vector, mCherry-tagged ScRce1, or mCherry-tagged LI0758, all under the control of the GAL1 promoter. These strains were grown in SC-Leu-Ura medium, diluted to a 1:10 ratio, and induced for expression over 8 h at 30 ℃. Live-cell imaging was performed using fluorescence microscopy with appropriate filter settings. The scale bar is 10 μm.