Fig. 4

Oscillation of target site mutations during proliferation of the single-cell subclone 1C3-1 derived from 1C3. A Schematic of experimental outline. Proliferation of the single-cell clone 1C3-1 was divided the first culture cycle that continues for 30 days and the second culture cycle that continues after 1 year storage. Target site mutations were determined by targeted PCR amplicon deep sequencing of gDNA extracted from cells at different time points of cell proliferation as indicated. D0, day 0; D15, day 15; D30, day 30; Mon12, year 1; Mon15, year 1 + 3 months; Mon12, year 1 + 6 months. B Changes in target site mutation profiles of 1C3-1 at indicated time points of cell proliferation (i.e., days 0, 15, and 30 and Mon12, Mon15, and Mon18). Gene targets and their mutation types are shown on top and at bottom, respectively. The numbers flanking Del or Ins indicated at bottom are the numbers of deleted or inserted nucleotides. √Frequency representing square root of mutation frequencies is indicated from the highest 1 in dark green to the lowest 0 in white. The top line chart and the boxplot under show the number of single-cell clones with a specific mutation type, i.e., number of clones, and the percentage distribution of a specific mutation type in all single-cell clones, i.e., proportion (%), respectively. C–F Representative target mutation oscillation of Rb1 (C), Lkb1 (D), Arid1a (E), and Smad4 (F) during cell proliferation of 1C3-1