Intrinsic checkpoint deficiency during cell cycle re-entry from quiescence

Jacob Peter Matson; Amy M. House; Gavin D. Grant; Huaitong Wu; Joanna Perez; Jeanette Gowen Cook

mark standard deviation (SD), n=3 biological replicates. WT and p53 KO cells were compared by unpaired, two tailed t test. SiCdt1 A p=0.0122*, siCdt1 B p=0.0011**. e. Percentage of underlicensed cells from Fig. 4C . Horizontal bars indicate means, error bars mark standard deviation (SD), n=3 biological replicates. WT and p53 KO cells were compared by unpaired, two tailed t test. SiControl p=0.0541 (ns), siCdt1 A p=0.0028**, siCdt1 B p=0.0011** f. Mean length of G1 (green), S (purple), and G2/M (grey) phases in hours for cells treated as Fig. 4C . G1 length: WT + siControl=10.9 hrs, WT + siCdt1 A = 25.7 hrs, WT + siCdt1 B= 35.5 hrs, p53 KO + siControl=14.1 hrs, p53 KO + siCdt1 A=7.6 hrs, p53 KO + siCdt1 B=16 hrs. Error bars are SD, n= minimum 3 biological replicates. Cell cycle phase percentages from DNA Content vs DNA synthesis were multiplied by the doubling time in hours to calculate hours for each phase. Doubling time is the mean of 3 biological replicates. The copyright holder for this preprint (which was not peer-reviewed) is the author/funder. It . https://doi.org/10.1101/558783 doi: bioRxiv preprint Figure 6 . Cdc6 dynamics in the first and second cycles after G0. a. RPE1-hTert cells constitutively expressing Cdc6-mVenus, PCNA mTurq2 and a CDK activity sensor (DHB-mCherry), and synchronized in G0 by contact inhibition and release were imaged every 10 minutes for 72 hours. Representative micrographs are shown marking the time of Cdc6 accumulation, CDK activity increase and S phase entry. Images were brightness/contrast adjusted. Representative cell from n=50 analyzed cells. The scale bar is 10 µm and applies to all images.