Fig 1.
Cell desynchronization via double thymidine block and release.
a) Cell cycle phases as indicated by cell DNA content and approximate phase distribution in an asynchronous population. b) Fluorescent profile of propidium iodide (PI) stained cells during asynchronous growth from t = 0 to t = 88. c) Fluorescent profile of PI-stained cells following G1/S synchronization by double thymidine block from t = 0 to t = 88. d) Percentages of cells in a given cell cycle phase at a given time point; asynchronous cell growth in green and desynchronous cell growth in red. The cell cycle phase percentages for each time point were determined via the Dean-Jett-Fox model.
Fig 2.
Rate of desynchronization using Kuiper test statistic.
a) Pairwise comparison of PI CDFs for each time point (data shown is from synchronized cells). b) Visual representation of Kuiper Test Statistic determination between time points. c) Rate of desynchronization between asynchronous (green) and synchronized (red) Hela cells. Over time (~60 hours) synchronized cells being to reach an asynchronous state.
Fig 3.
Single cell model of desynchronization.
a) DNA synthesis is captured by the Gaussian error function where the relative durations of cycle phase are tunable. b) Simulated data of PI staining of multiple lineages with normally distributed initial gene content. c) Cell cycle pace inheritance following a Gaussian distribution. d) Desynchronization rate of simulated cell population.
Fig 4.
Noise variation of cell periodicity.
a) Representative images of time lapse experiments. 100 cells were tracked for each condition and the population mean and standard deviation of cell cycle duration was determined. Once the septum (white arrow) is visible following cytokinesis, the cell cycle duration recording begins for both daughter cells (yellow and blue arrow). Both cells being cell cycle at Frame 2, and both daughter cells can be seen progressing through interphase in Frames 26–28. By the end of Frame 57, the first daughter cell completes the cell cycle and recording ends. The second daughter cell (yellow arrow) had a substantially longer cell cycle duration, which concluded at the end of Frame 84, thus demonstrating the inherent variability of cell cycle duration between identical cells within the population. b) Asynchronous cells were wither treated with 1 μg/mL of LPS or left untreated and cell cycle duration was recorded (n = 100). c) Values obtained from time lapse microscopy for cell cycle mean and standard deviation were used in our model to predict the impact on cell cycle desynchronization. The model revealed the LPS administration should result in an increased rate of cell cycle desynchronization d) Cell cycle phase distribution of LPS treated cells following cell cycle synchronization for 88 hours post release (n = 3). e) Normalized ASF scores for LPS-treated desynchronizing cells. The asynchronous population was not normalized in order to capture the overall linear trend (n = 3).