Fig. 4: High-resolution iFlpMosaics reveal the impact of genetic mutations on single-cell clonal dynamics.

a, Quantification of Ki67⁺ cells (in cycle) and EdU⁺ cells (S phase) in livers. b, Left: a schematic of the alleles and how they can be used to fluorescently barcode and track the clonal expansion of single mutant/MTomato⁺ and wild-type (WT)/MYFP⁺ cells from P1 to P20. Right: representative confocal micrographs with magnified insets (white boxes) of P20 livers (from animals induced with 4-OHT at P1) with different recombination frequencies (Extended Data Fig. 6). c, The clone size distributions of hepatocytes at both low and high recombination rates. d, The whole-liver weight at P1 and P20. e, A quantification of MYFP⁺ (Myc^WT) and MTomato⁺ (Myc^KO) clone size frequency. f, A chart depicting the normalized expansion of Myc^WT and Myc^KO hepatocytes. g, The representative confocal micrographs of P20 pancreas from control and Myc-floxed mice carrying the indicated iFlpMosaic alleles. Right: inverted LUT images depicting the clone mapping (Fiji image analysis scripts) according to their dual color code. h, The quantification of clone size frequency. i, A chart depicting the normalized expansion of Myc^WT and Myc^KO pancreatic cells. j, log₂ ratio of MTomato⁺ cells to MYFP⁺ cells in control and Myc-floxed pancreas, showing a 3.7-fold decrease of MTomato⁺ (Myc^KO) cells over 20 days. k, Representative confocal micrographs of adult livers from control and Myc-floxed mice in which the indicated alleles were induced 14 days before tissue collection. Right: a semiautomatic mapping of clones according to their fluorescent barcode. Note the loss of MTomato⁺ cells in the Myc-floxed background. l, The clone size, showing that WT hepatocytes rarely divide or expand over a 2 week period. m, The clone frequency, showing the lack of impact of Cre expression in WT MTomato⁺ cells. n, The percentages of proliferative (composed of more than one cell) and nonproliferative clones. o, log₂ ratio of the MTomato⁺ and MYFP⁺ cell frequency in control and adult Myc-floxed livers. p, The clone frequency Myc^WT and Myc^KO hepatocytes. q, The percentages of proliferative and nonproliferative Myc^WT and Myc^KO clones. The data are presented as mean values ± standard deviation. Scale bars, 100 μm.

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