Supplementary Figure 1: Representative iCC Confetti analysis and Confetti-based estimate fidelity.
From: Lifelong haematopoiesis is established by hundreds of precursors throughout mammalian ontogeny
(a) CFP, GFP, YFP or mCherry positive HL60 cells were used as single color controls during flow cytometry Confetti analysis. (b) Untreated and 4-OHT treated iCC cultures are shown to illustrate gating strategy for distinct Confetti colors. (c) Results of computer simulation of variance in the proportion of Confetti colors across a range of starting cell numbers (10 to 10,000 cells) and a range of starting percentages of a given Confetti color. The Log10(CV) vs. Log10(cell number) is shown. Simulated data was fixed at each percentage and sampled 15,000 times for each sample size of each percentage. The resulting regression lines were stable with respect to slope (−0.5) across all percentages except the 10% simulation measured the minimum number of times (top line). (d,e) Immortalized fibroblasts, like iCCs, are susceptible to polyploidy or chromosomal duplication during in vitro culture. Thus, a subset of cells acquire extra Confetti alleles and can be labeled with multiple Confetti colors after exposure to 4-OHT (e. g. RFP + CFP + cells in b). (d) About 2% of iCCs displayed >4N DNA content. (e) Karyotype confirms presence of polyploid iCCs.
