Figure S3.
Analysis of quiescence entry after cell cycle synchronization or nutrient depletion and linear discriminant analysis of cluster separation by stress factors.(A) A two-step k means–based algorithm to sort time series before TPML. Heatmaps = single-cell time series derived from 6C1 cells that were M-phase (nocodazole) synchronized before starvation. Clustering identifies cells that exited from G1 shortly after starvation onset. (B) Average time series for cell cycle (Cdc10 signal) and stress markers (Sfp1, Rtg1, Gln3, Stb3, and Xbp1) in M-phase arrest-released 6C1 cells that exited G1 shortly before starvation onset (n = 3; 96 cells, OAM425). (C) Average time series for cell cycle (Cdc10 signal) and stress markers (Sfp1, Rtg1, Gln3, Stb3, and Xbp1) in G1-released 6C1 cells that exited G1 shortly before starvation onset (n = 3; 86 cells, OAM425). (D) Average time series of cell cycle (Cdc10 signal) and stress markers (Sfp1, Rtg1, Gln3, Stb3, and Xbp1) in 6C1 cells that were exposed to glucose depletion for 2 h before quiescence entry (n = 3; 504 cells, OAM425). (E) Average time series of cell cycle (Cdc10 signal) and stress markers (Sfp1, Rtg1, Gln3, Stb3, and Xbp1) in 6C1 cells that were exposed to nitrogen depletion for 2 h before quiescence entry (n = 3; 222 cells, OAM425). (F) Average cluster separation at each time point between low- and high-Cdk1 Q-cells, as measured by the Mahalanobis distance calculated using LDCs based on the time series for each stress marker derived from OAM425 (Fig. 3) and OAM667 (Fig. 4) cells. (G) Average probability of high-Cdk1 quiescence assigned to high-Cdk1 Q-cells by SVMs based on final cell size after rich-starvation medium transition. Solid lines with shaded area = average ± 95% confidence intervals from biological replicates. Green arrowhead = onset of nutrient depletion. X-axis red arrowheads = onset of starvation. Y-axis red arrowhead with dotted line = a priori probability.

Analysis of quiescence entry after cell cycle synchronization or nutrient depletion and linear discriminant analysis of cluster separation by stress factors. (A) A two-step k means–based algorithm to sort time series before TPML. Heatmaps = single-cell time series derived from 6C1 cells that were M-phase (nocodazole) synchronized before starvation. Clustering identifies cells that exited from G1 shortly after starvation onset. (B) Average time series for cell cycle (Cdc10 signal) and stress markers (Sfp1, Rtg1, Gln3, Stb3, and Xbp1) in M-phase arrest-released 6C1 cells that exited G1 shortly before starvation onset (n = 3; 96 cells, OAM425). (C) Average time series for cell cycle (Cdc10 signal) and stress markers (Sfp1, Rtg1, Gln3, Stb3, and Xbp1) in G1-released 6C1 cells that exited G1 shortly before starvation onset (n = 3; 86 cells, OAM425). (D) Average time series of cell cycle (Cdc10 signal) and stress markers (Sfp1, Rtg1, Gln3, Stb3, and Xbp1) in 6C1 cells that were exposed to glucose depletion for 2 h before quiescence entry (n = 3; 504 cells, OAM425). (E) Average time series of cell cycle (Cdc10 signal) and stress markers (Sfp1, Rtg1, Gln3, Stb3, and Xbp1) in 6C1 cells that were exposed to nitrogen depletion for 2 h before quiescence entry (n = 3; 222 cells, OAM425). (F) Average cluster separation at each time point between low- and high-Cdk1 Q-cells, as measured by the Mahalanobis distance calculated using LDCs based on the time series for each stress marker derived from OAM425 (Fig. 3) and OAM667 (Fig. 4) cells. (G) Average probability of high-Cdk1 quiescence assigned to high-Cdk1 Q-cells by SVMs based on final cell size after rich-starvation medium transition. Solid lines with shaded area = average ± 95% confidence intervals from biological replicates. Green arrowhead = onset of nutrient depletion. X-axis red arrowheads = onset of starvation. Y-axis red arrowhead with dotted line = a priori probability.

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