Time-lapse images (A) and representative single-cell traces (B) of p53-Venus level in MCF7 cells following 1 etoposide treatment. μM Figure S2. Dynamic features and accumulation time combinatorially determine cell fates. (A and B) Dose and duration-dependency of mild (A) and acute (B) Dox-induced cellular apoptosis. DNA content of cells was measured using PI staining by flow cytometry at 1st~4th day after a mild Dox treatment and 12~48 hrs after an acute Dox treatment. Cellular apoptosis was also measured using Annexin V/DAPI staining at the ending time point. Control cells were (C) cultured in drug-free medium (0.1% DMSO) and collected for analysis at indicated time. Western blot analysis of BAX and PUMA after the mild and acute Dox-treatment at indicated time. Control cells were cultured in drug-free medium (0.1% DMSO) for 24 hrs. The immunoblotted intensities of each protein compared to GAPDH were quantified by densitometry and expressed in relative fold change to the control group. Figure S3. E∫p53, rather than pulse number or controls cell fate. ∫p53, (A-C) Pulse amplitude (A), lifespan (B) and terminal amplitude (C) held constant across cells that exhibited terminal pulse under the treatment of 0.01, 0.05 and 0.1 Dox within 96 hrs. μM Number of cells (n) = 121 (0.01 n= 127 (0.05 and n= 118 (0.1 μM), μM) μM). The average pulse amplitude (D), pulse duration (E), integrated p53 (∫p53) level (F) and (D-G) pulse number (G) before terminal pulse in apoptotic (n= 227) and survival (n= 139) cells.
Time-lapse images (A) and representative single-cell traces (B) of p53-Venus level in MCF7 cells following 1 μM etoposide treatment. Figure S2. Dynamic features and accumulation time combinatorially determine cell fates. (A and B) Dose and duration-dependency of mild (A) and acute (B) Dox-induced cellular apoptosis. DNA content of cells was measured using PI staining by flow cytometry at 1st~4th day after a mild Dox treatment and 12~48 hrs after an acute Dox treatment. Cellular apoptosis was also measured using Annexin V/DAPI staining at the ending time point. Control cells were cultured in drug-free medium (0.1% DMSO) and collected for analysis at indicated time. (C) Western blot analysis of BAX and PUMA after the mild and acute Dox-treatment at indicated time. Control cells were cultured in drug-free medium (0.1% DMSO) for 24 hrs. The immunoblotted intensities of each protein compared to GAPDH were quantified by densitometry and expressed in relative fold change to the control group. Figure S3. E∫p53, rather than pulse number or ∫p53, controls cell fate. (A-C) Pulse amplitude (A), lifespan (B) and terminal amplitude (C) held constant across cells that exhibited terminal pulse under the treatment of 0.01, 0.05 and 0.1 μM Dox within 96 hrs. Number of cells (n) = 121 (0.01 μM), n= 127 (0.05 μM) and n= 118 (0.1 μM). (D-G) The average pulse amplitude (D), pulse duration (E), integrated p53 (∫p53) level (F) and pulse number (G) before terminal pulse in apoptotic (n= 227) and survival (n= 139) cells.
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