A role for cell polarity in lifespan and mitochondrial quality control in the budding yeast Saccharomyces cerevisiae

Enrichment of aging yeast cells and budding polarity assay in Saccharomyces cerevisiae

“…The efficacy of the mCAD can be verified by determining the age, size, and viability of isolated cells. Biotinylated progenitor cells isolated from the mCAD exhibit aging phenotypes including increased abundance of bud scars, increased cell size and decreased cell viability, as a function of the time of propagation in the mCAD ( Figures 7 A–7C) ( Yang et al., 2022 ). The viability of yeast cells can be determined by trypan blue, methylene blue, propidium iodide/fluorescein diacetate, or LIVE/DEAD staining and counted by hemacytometer ( Figure 6 ) ( Kwolek-Mirek and Zadrag-Tecza, 2014 ).…”

“…The viability of yeast cells can be determined by trypan blue, methylene blue, propidium iodide/fluorescein diacetate, or LIVE/DEAD staining and counted by hemacytometer ( Figure 6 ) ( Kwolek-Mirek and Zadrag-Tecza, 2014 ). WGA-AlexaFlour or Calcofluor White can be used to assess buds scars and replicative aging or age-associated declines in polarity ( Figure 7 E) ( Yang et al., 2022 ). Isolated cells can be analyzed by light microscopy ( Sing et al., 2022 ; Yang et al., 2022 ) as well as transcriptomic, proteomic or epigenetic approaches ( Hendrickson et al., 2018 ).…”

“…WGA-AlexaFlour or Calcofluor White can be used to assess buds scars and replicative aging or age-associated declines in polarity ( Figure 7 E) ( Yang et al., 2022 ). Isolated cells can be analyzed by light microscopy ( Sing et al., 2022 ; Yang et al., 2022 ) as well as transcriptomic, proteomic or epigenetic approaches ( Hendrickson et al., 2018 ).

Figure 6 Methods to determine cell viability Cells can be stained by trypan blue, methylene blue, propidium iodide + fluorescein diacetate, or LIVE/DEAD dye.

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“…We also describe an approach to assess bud site selection, a marker for cell polarity, during the aging process. For complete details on the use and execution of this protocol, please refer to Yang et al. (2022) .…”

“…For complete details on the use and execution of this protocol, please refer to Yang et al. (2022) .…”