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. 2018 Dec 1;32(23-24):1499–1513. doi: 10.1101/gad.318485.118

Figure 1.

Figure 1.

Analysis of individual telomerase-deficient lineages reveals frequent prolonged nonterminal arrests. (A) Sequential images of telomerase-negative (TetO2-TLC1) cells trapped in a microcavity of the microfluidic device. Hours spent after addition of 30 µg/mL dox to repress telomerase activity are shown in yellow. (B) Display of consecutive cell cycle durations of TetO2-TLC1 lineages grown in the microfluidic device as in A (n = 187, 40 of which were already published in our previous work) (Xu et al. 2015). Cells were monitored overnight before (−dox) and then for successive generations after (+dox) addition of 30 µg/mL dox to inactivate telomerase (designated generation 0). Each horizontal line is an individual cell lineage, and each segment is a cell cycle. Cell cycle duration (in minutes) is indicated by the color bar. X at the end of the lineage indicates cell death, whereas an ellipsis (…) indicates that the cell was alive at the end of the experiment. (Inset) Magnified view of several lineages showing nonterminal arrests (black). (C) Distribution of all cell cycle durations taken from telomerase-negative (red; n = 5962) and telomerase-positive (black; n = 1895) lineages shown in B and Supplemental Figure S1. Percentages indicate the fraction of cell cycles >150 min (first vertical black line) or >360 min (second vertical black line) for each lineage. (D) Distribution of nonterminal cell cycle durations as a function of generation for telomerase-negative (left; n = 5775) and telomerase-positive (right, n = 1887) cells extracted from B and Supplemental Figure S1. The color bar indicates the frequency. (E) Frequency of all nonterminal arrests (cell cycle >150 min; blue circles) and prolonged nonterminal arrests (cell cycle >360 min; red triangles) extracted from B and Supplemental Figure S1 as a function of generation for telomerase-negative (left) and telomerase-positive (right) lineages. The gray-shaded area encompasses points based on less than five lineages, for which the data may be less reliable. See also Supplemental Figure S1.