Rapid Nuclear Exclusion of Hcm1 in Aging Saccharomyces cerevisiae Leads to Vacuolar Alkalization and Replicative Senescence

Supplemental Material for Ghavidel et al., 2018

• Figure S1 -

  (A)Live cell imaging of dividing WT yeast.  (B) Live cell imaging of dividing yeast co-stained with the vacuolar membrane marker FM4-64 and calcofluor-white. (C) cDCFDA fluorescence emission spectra of young and old yeast. (D)Live cell imaging of dividing Matα yeast cells expressing either Ste3-GFP, or the GFP variant pHluorin fused to the
  pheromone receptor Ste3. (.pptx)
  

• Figure S2 -

  (A)Expression ratios of RT-PCR RNA expression analyses of vacuolar proton pump subunits (VMA) and a vacuolar fusion factor (YPT7) in replicatively old cells enriched using MEP.  (B) CMAC-Arg fluorescence emission spectra of cells in (Fig. 2B) (n=25).(.pptx)
  

• Figure S3 -

  (A) Live cell microscopy of WT yeast expressing a genomically integrated gal-inducible α-syn-GFP construct. (B) RNA expression analysis of autophagosome (ATG) subunits in early log (young) cells and a population enriched for replicatively old cells using MEP. (.pptx)
  

• Figure S4 -

  Consensus binding motifs for Forkheadtranscription factors in the 5’ upstream regions of VMA genes (http://yetfasco.ccbr.utoronto.ca). (.pptx)
  

• Figure S5 -

  Additional images of aging cells expressing endogenous GFP C-terminal fusions to the FKH1, FKH2 and HCM1 genes. (.pptx)
  

• File S1 -

  Contains Tables S1 - S5. (.pdf)