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. 2021 May 13;10:e65234. doi: 10.7554/eLife.65234

Figure 2. Variable cell size and clonal outgrowth in post-cisplatin-treated cells.

(A) Representative images from cells treated as per Figure 1A showing control (Cont.), proliferative (Prolif.), and arrested (Arrest) cells. Scale bars = 100 µm. (B) Cell and nuclear size were calculated on sub-populations of cells that appeared to form clonal outgrowths. Quantification of cell size and nuclear size from (A), with a minimum of n = 100 cells analysed per condition. Shown are the mean ± SD. Statistical significance was determined by one-way ANOVA (****p<0.0001, n.s = not significant). (C) LeGO RGB colour-guided clonal cell tracking system was used to track clonal dynamics after cisplatin pulse treatment. Cells were treated as per Figure 1A, with clonal identification and quantification (D), measured at 3 and 21 days post-cisplatin exposure using Image J/Fiji (42 days post-cisplatin exposure for NCI-H1573).

Figure 2.

Figure 2—figure supplement 1. Senescence and LeGo Clonal Recovery of LUAD cells following cisplatin pulse.

Figure 2—figure supplement 1.

(A) Representative images and quantification of A549, NCI-H1573, and CI-H1299 cells 3 days after cisplatin exposure stained for beta-galactosidase. Scale bar = 100 µm. Shown are the mean ± SD of three biological repeats. Statistical significance was determined by unpaired two-tailed Student’s t-test (****p<0.0001, ***p<0.001, **p<0.01). (B) FACS analysis of beta-galactosidase activity levels (C12FDG) versus ‘cell size’ (forward scatter [FSC]) and (C) p21, p16 and at 3 days after cisplatin exposure. Shown are the mean ± SD of three biological repeats. Statistical significance was determined by unpaired two-tailed Student’s t-test (****p<0.0001, ***p<0.001, *p<0.05). (D) Three biological repeat experiments of LeGO RGB colour-guided clonal cell tracking system were used to track clonal dynamics after cisplatin pulse treatment in each cell line. Cells were treated as per Figure 1A, with clonal identification and quantification measured at 3 and 21 days post-cisplatin exposure using Image J/Fiji (42 days post-cisplatin exposure for NCI-H1573). For summary data, only colours that represented >0.1% of total population were scored.