Fig. 3
From: Clonal dynamics in osteosarcoma defined by RGB marking
RAINBONE tumours showed dominant clones in the long term. a Representative image of tumours 50 days after RAINBONE inoculation (n = 14). 1, Primary implantation site (HA/TCP ceramic region) showing polyclonal evolution; 2, transition zone; 3, large monoclonal areas generated outside of the ceramic region (extracompartmental region). b For comparison, representative image of tumours at 25 days, with tumour growth starting in the extracompartmental region. c LAM-PCR analysis showing the rich amplification of different provirus integration sites in both short- and long-term RAINBONE tumours. Negative control = water. d Representative detailed image of tumour heterogeneity present in the large clonal area. Low-frequency clones are indicated by white arrows. e Representative SPADE tree clonal composition of RAINBONE tumours at 50 days. Note that in this case, colours represent the population frequency. f Representative 3D flow cytometric RAINBONE tumour analysis at 50 days (n = 4). Arrows indicate the most frequent clones that were subsequently FACS sorted (right). g LAM-PCR analysis showing the monoclonal origin of FACS sorted clones (n = 3). White bar = 200 µm; orange bar = 100 µm; blue bar = 50 µm
