Volume growth in animal cells is cell cycle dependent and shows additive fluctuations

The way proliferating animal cells coordinate the growth of their mass, volume, and other relevant size parameters is a long-standing question in biology. Studies focusing on cell mass have identified patterns of mass growth as a function of time and cell cycle phase, but little is known about volume growth. To address this question, we improved our fluorescence exclusion method of volume measurement (FXm) and obtained 1700 single-cell volume growth trajectories of HeLa cells. We find that, during most of the cell cycle, volume growth is close to exponential and proceeds at a higher rate in S-G2 than in G1. Comparing the data with a mathematical model, we establish that the cell-to-cell variability in volume growth arises from constant-amplitude fluctuations in volume steps rather than fluctuations of the underlying specific growth rate. We hypothesize that such ‘additive noise’ could emerge from the processes that regulate volume adaptation to biophysical cues, such as tension or osmotic pressure.


Sample-size estimation
 You should state whether an appropriate sample size was computed when the study was being designed  You should state the statistical method of sample size computation and any required assumptions  If no explicit power analysis was used, you should describe how you decided what sample (replicate) size (number) to use Please outline where this information can be found within the submission (e.g., sections or figure legends), or explain why this information doesn't apply to your submission:

Replicates
 You should report how often each experiment was performed  You should include a definition of biological versus technical replication  The data obtained should be provided and sufficient information should be provided to indicate the number of independent biological and/or technical replicates  If you encountered any outliers, you should describe how these were handled  Criteria for exclusion/inclusion of data should be clearly stated  High-throughput sequence data should be uploaded before submission, with a private link for reviewers provided (these are available from both GEO and ArrayExpress) Please outline where this information can be found within the submission (e.g., sections or figure legends), or explain why this information doesn't apply to your submission: No power analysis was used. All the analyses presented here use a dataset of 1696 single cell volume growth curves acquired over 4 distinct experiments. This sample size is very high compared to what most other available techniques have previously reported and thus improves the statistical resolution of animal cell growth analysis. For each plot, the figure legend specifies the minimum number of individual cells per bin used for the analysis.
Four independent experiments were performed to constitute this dataset. The protocol was the same (see Methods section) for all replicates and we checked that the replicates showed a similar growth rate before combining them ( Figure  1 -Supplement 1). The visual curation and algorithm used to remove the few outliers corresponding to errors in the automated image segmentation and tracking are described in the Methods Section. Following this step, no cell was excluded from the analysis.

Statistical reporting
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Please outline where this information can be found within the submission (e.g., sections or figure legends), or explain why this information doesn't apply to your submission: (For large datasets, or papers with a very large number of statistical tests, you may upload a single table file with tests, Ns, etc., with reference to sections in the manuscript.)

Group allocation
 Indicate how samples were allocated into experimental groups (in the case of clinical studies, please specify allocation to treatment method); if randomization was used, please also state if restricted randomization was applied  Indicate if masking was used during group allocation, data collection and/or data analysis Please outline where this information can be found within the submission (e.g., sections or figure legends), or explain why this information doesn't apply to your submission: Additional data files ("source data")  We encourage you to upload relevant additional data files, such as numerical data that are represented as a graph in a figure, or as a summary table  Where provided, these should be in the most useful format, and they can be uploaded as "Source data" files linked to a main figure or table  Include model definition files including the full list of parameters used  Include code used for data analysis (e.g., R, MatLab)  Avoid stating that data files are "available upon request" Please indicate the figures or tables for which source data files have been provided: All statistical information can be found in the figures and figure legends.
The four experiments were performed identically and either all grouped together or shown individually to show the reproducibility of the trend across replicates (Figure 2a and Figure 3).
The dataset, R code for the analysis and C code for the simulations are all available on the following repository: https://datadryad.org/stash/share/57Obey5DkCwX393xddneQI-Td4vCAQUhbbik2lUZRLA which is available to the reviewers and will be made public when the article is accepted for publication.