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. 2022 Jul 28;13:918–943. doi: 10.18632/oncotarget.28256

Figure 8. The gph1Δ-dependent increase in glycogen concentration does not affect processes 1, 2, 3 and 4 either in yeast cultured under caloric restriction (CR) conditions without LCA or in yeast cultured under non-CR conditions with lithocholic acid (LCA).

Figure 8

Wild-type (WT) and gph1Δ cells were cultured in the nutrient-rich YP medium initially containing 0.2% glucose (CR conditions) or 2% glucose (non-CR conditions) with LCA. Aliquots of differently aged cell populations were recovered from the logarithmic (L), diauxic (D), post-diauxic (PD) or stationary (ST) growth phase. High-density quiescent (Q) cells and low-density Q cells were purified from these cell populations with the help of the centrifugation in Percoll density gradient, as described in Materials and Methods. (A and B) Mean diameters of high-density Q cells recovered from different growth phases are shown for yeast cultured under CR conditions without LCA (A) and yeast cultured under non-CR conditions with LCA (B). Mean diameters of high-density Q cells in A and B were measured using differential interference contrast microscopy and subsequent morphometric analysis, as described in Materials and Methods. (C and D) The values of the percentage of high-density Q cells in each of the differently aged cell populations are shown for yeast cultured under CR conditions without LCA (C) and yeast cultured under non-CR conditions with LCA (D). The percentage of high-density Q cells in C and D was measured with the help of a cell counter, as described in Materials and Methods. (E–H) The clonogenicities of low-density (E, F) and high-density (G, H) cells in each of the differently aged cell populations are shown for yeast cultured under CR conditions without LCA (E, G) and yeast cultured under non-CR conditions with LCA (F, H). The clonogenicities of low- and high-density cells were assessed using a plating assay for reproductive (colony forming) capability, as described in Materials and Methods. Other Abbreviations: HD: high-density cells; LD: low-density cells.