Fig. 3

Barcode-mediated clonal tracking of metastasis in vivo. a A schematic depiction of the experimental design used for introducing the lentiviral barcode library into freshly isolated PDX tumor cells is shown. Barcoded tumors were engrafted into the MFPs of four recipient mice denoted A, B, C, and D. Due to the high complexity of the barcode library, each mouse was engrafted with cells harboring non-overlapping barcodes. b A schematic representation of barcoded MFP tumors and metastatic tumors that were isolated from each mouse for barcode sequencing to monitor clonal dynamics during metastasis. Lu (lung), Li (liver), Br (brain). c Unique dominant barcodes (the top 95% most abundant) in each sample were quantified and plotted. The mean quantity of unique barcodes present in each organ site is depicted above each bar. (two-tailed T-tests, *p = 6.285e−9, **p = 1.830e−5, ***p = 0.0170; error bars are standard error of the mean, SEM; n = 4 MFP tumors, n = 21 lung, n = 12 liver, and n = 4 brain metastases). d Shannon Diversity Indices were calculated (in nats: natural digits) taking into account all barcodes for each sample as a measure of ITH. (two-tailed T-tests, *p = 9.050e−15, **p = 3.124e−9, ***p = 7.581e−3, error bars are SEM; n = 4 MFP tumors, n = 21 lung, n = 12 liver, and n = 4 brain metastases), ref (reference sample). e The percent of dominant barcodes (top 95%) in each metastatic lesion relative to the total number of unique barcodes detected in its matched MFP tumor is shown. f The distribution (fraction of total barcode reads) of dominant barcodes (top 95%) in each tumor sample from mouse C is shown. CPM (counts per million)