Extended Data Fig. 6: Mapping the clonal expansion of single mutant and wildtype cells with iFlpMosaics.
a,b, Representative confocal images (low magnification large tile scans) of entire P20 liver sections from mice carrying the indicated alleles. Recombination of the reporter alleles was induced at P1 and resulted in different recombination frequencies. Right panels show the results of automatic image processing with Fiji scripts, which generate spatial maps of dual-labeled clones of cells (with membrane and nuclear labeling). The bigger the circle, the bigger the clone. c, Quantification and representative confocal micrographs of proliferative pancreatic cells (Ki67+ in cycle or EdU+ in S-phase) at different postnatal stages. d, Quantification and representative confocal micrograph of Ki67+/DAPI+ cells in the adult liver. e, Representative confocal micrographs of adult liver sections from control and Myc-floxed mice carrying the indicated alleles. Left panels show endogenous fluorescent signals from the reporter alleles. Center panels show sections from the same mice immunostained with antibodies to DsRed (detects MTomato and H2B-Cherry), GFP (detects MYFP and H2B-GFP), or the HA epitope (detects MTFP1 and H2B-Cerulean) together with Fiji-script-generated spatial maps of dual-labeled clones (with membrane and nuclear labeling) reveal hepatocyte ploidy and clone size. Graphs show the percentages of mononucleated cells, binucleated cells, and 2-cell mononuclear clones. In control mice, MYFP+ (WT) and MTomato+ (WT) diploid cells occur in similar proportions, whereas in Myc-floxed mutants MTomato+ (MycKO) diploid cells are more frequently binucleated and give rise to fewer 2-cell mononucleated clones than MYFP+ (MycWT) cells. Scale bars, 50μm (c, d), the rest 300μm. Data are presented as mean values +/− SD. For numerical data see Source Data file 1.
