Figure 5. Clonal populations derived from sorted cells maintain the bi-stable phenotype of chl-roGFP response to H₂O₂.

(A–C) The distribution of chl-roGFP OxD (%) 40–45 min post 80 µM H₂O₂ treatment in clonal populations derived from sorted single cells of different origins, 3 weeks post sorting. The ‘reduced’ (B) and ‘oxidized’ (C) subpopulations were sorted 30 min post 80 µM H₂O₂ treatment based on chl-roGFP oxidation (Figure 3—figure supplement 1); ‘control’ (A) – clones of untreated cells that were sorted based on positive roGFP fluorescence. Each histogram is of a single clone,≥9900 cells per histogram, six representative clones per group are shown. (D) The fraction of dead cells 24 hr post H₂O₂ treatment of the different clones shown in (A–C) as measured by positive Sytox staining. Data is shown as mean ± SEM, n = 6 clones per group per treatment.

Figure 5—figure supplement 1. Sorted clonal populations maintain the bi-stable phenotype in chl-roGFP response to H₂O₂.

(A–B) The distribution of chl-roGFP OxD 40–45 min post re-exposure to 80 µM H₂O₂ treatment in clonal populations derived from sorted single cells of different origins, 3 weeks post sorting. The ‘reduced’ (A) and ‘oxidized’ (B) subpopulations were sorted 100 min post 80 µM H₂O₂ treatment based on chl-roGFP oxidation. Each histogram is of a single clone,≥9900 cells per histogram, six representative clones per group are shown. The same phenomenon was observed in two independent experiments in all examined clones (≥18 clones per group), for visualization data from one is shown. (C) The fraction of dead cells 24 hr post H₂O₂ treatment of clones originating from single cells sorted 30 and 100 min post H₂O₂ treatment or untreated control, as measured by positive Sytox staining. Data is shown as mean ± SEM, n ≥ 5 per column. Data of 2 independent experiments.