Autophagy Induction in Yeast

The experiment was conducted as previously described [14 (link)]. The YOM38 yeast strain containing pR316-GFP-ATG8 plasmid was incubated in the dark with YPD medium and shaking at 180 rpm. After 24 h, cells were cleaned with PBS, divided into different groups with the same OD₆₀₀ value and treated with GTS B at different concentrations of 0, 1, 3 and 10 µM and RES at 300 μM as positive control. Cells were cultured for 22 h, washed with PBS and stained with DAPI (4′, 6 diamidino-2-phenylindol 20 μg/mL) for 10 min in dark, and then the cells were cleaned with PBS to remove remaining DAPI from the background. Finally, induction of autophagy of yeast was observed using a two-photon confocal fluorescence microscope (Olympus FV1000BX-51, Tokyo, Japan).