Protein dynamics for proliferating and spontaneously quiescent MCF10A cells.

<p>(A–I) Column 1: Time-lapse imaging of CDK2 activity in asynchronous cells was followed by fixation and IF staining for the indicated protein. Protein signals were then reconstructed as a function of time-since-anaphase for CDK2^inc cells (blue dots) and CDK2^low cells (red dots), as in <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.2003268#pbio.2003268.g001" target="_blank">Fig 1H</a>. The protein signal in prolonged quiescent cells is plotted at the 24-hour mark (purple dots). The approximate time spent in G1, S, or G2 is marked above the plots, based on EdU incorporation data from <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.2003268#pbio.2003268.g001" target="_blank">Fig 1H</a>. Column 2: Moving average through the blue, red, or purple points from Column 1, as in <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.2003268#pbio.2003268.g001" target="_blank">Fig 1I</a>. Error bars represent standard deviation. Column 3: Protein signal as a function of time since CDK2 activity buildup (R-point), as in <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.2003268#pbio.2003268.g001" target="_blank">Fig 1J</a>. The y-axis units are arbitrary fluorescence units. Total number of cells plotted: (A) 1,949; (B) 433; (C) 1,549; (D) 1,729; (E) 2,596; (F) 1,043; (G) 1,885; (H) 2,117; (I) 1,300. The data for each antibody are pooled from 8 replicate wells (1 image per well). Abbreviations: CDK2, Cyclin-Dependent Kinase 2; IF, immunofluorescence; R-point, Restriction Point.