Fig. 2: In vivo cellular barcoding using the LoxCode mouse line.
a Schematic of the LoxCode construct: LoxP sites (arrowheads), 1–13: unique DNA sequences (elements). LoxCode construct size = 616 nucleotides. b Representative examples of LoxCode construct PCR products before and after recombination with Rosa26 or Cdh5 driven ERT2cre recombinase. Number of elements per fragment is indicated. Data is representative of n = 104 independent libraries prepared from crosses with Cdh5ERT2Cre mice and n = 180 independent libraries prepared from crosses with Rosa26ERT2Cre mice. c Proportionality of barcode amplification in control pools. (i) 1 element and 13 element barcodes are over- and under- represented respectively. (ii) 5–9 element barcode sequences are within a near-linear manner, allowing for reliable quantification of relative clonal output to all assessed lineages (biomass). Note, multiple representation of 1 and 5 element (E) barcodes reflects the presence of multiple independent 1 and 5 element barcode samples in the experiment. Data shown derive from two independent experiments that were performed in technical duplicate. All data points are shown. r2 represents Pearson’s correlation coefficient for each independent experiment. d Complex barcodes (requiring more recombination steps) are rarer and therefore more likely to be clonal. Number of embryos in which barcodes of a given complexity (minimal number of recombination steps) are detected. n = 11 independent embryos. e % of quantifiable (complex 5–9 element) barcodes after 4-OHT injections. 1 h: 8 embryos; 6 h: 10 embryos, 12 h: 14 embryos, 24 h: 10 embryos, 48 h: 8 embryos, 120 h: 8 embryos. Data were analysed using One-way ANOVA (using Tukey’s P value adjustment) was used for multiple comparisons. Exact p values are shown. Bars represent mean ± SD.
