S. cerevisiae enters heterogeneous quiescent states under acute starvation. (A) WT cell cycle duration (strain OAM128) measured by the time between two consecutive cytokinesis events in rich medium (n = 13; 400 cells) or after exposure to acute starvation (n = 9; 300 cells). (B) Daughter/Mother cell size ratio at cytokinesis, measured as cell area in pixels, in rich medium (n = 14; 400 cells) or for cells that completed cell cycle under starvation (n = 9; 300 cells). (C and D) Representative phase-contrast (PC) micrographs for cell division in rich (C) and starvation (D) medium. (E) Scaled mean cellular fluorescence intensity of the trehalose metabolism marker Tps1 tagged with mNeonGreen (mNG; n = 9; 499 cells, OAM394) and the glycogen metabolism marker Gsy2-mNeonGreen (n = 9; 300 cells, OAM396) during starvation. (F) Population budding index during starvation (blue) and viability (green) of Q-cells upon return to proliferation (n = 3; 237 cells). (G) Representative PC micrographs of cells during the starvation–proliferation transition showing a budded Q-cell (yellow arrow). (H and I) Percentage of budded and unbudded Q-cells (H) after 24-h starvation (n = 6; 400 cells) and their viability measured as proliferation resumption upon return to rich medium (I). (J) Average cell volume of quiescent budded and unbudded cells during starvation. (K) Average Tps1-mNeonGreen intensity in budded and unbudded Q-cells. (L) Average Gsy2-mNeonGreen intensity in budded and unbudded Q-cells. All data from biological replicates. Red star = P < 0.05, K-S test. Solid lines with shaded area = average plus 95% confidence interval. Box plots: central mark, median; box bottom and top limit, 25th and 75th percentiles; whiskers, most extreme nonoutlier values. Approx., approximate.