Fig. 6

Single cell cloning and lineage tracing reveal that both lineage-restricted and bipotent progenitors are present within the PIK3CA^H1047R-iHBEC^CD117 cell line. A Representative FACS plots illustrating the gating strategy for the generation of single cell (sc) clones from the luminal progenitor cell line. TROP2⁺/CD271⁻ cells were sorted (left) as single cells and expanded. Note the presence of both bipotent (3D9) and lineage restricted (2D10) clones. B Multicolor live cell imaging of PIK3CA^H1047R-iHBEC^CD117 progenitor cells at clonal density expressing a K19-tagged miRFP670 (red) and K14 promoter-driven RFP reporter (green). Colonies were traced during ground state culture (left) and after differentiation (right). Two representative colonies are shown as a demonstration of K14⁺ cells emerging from K19⁺ luminal cells with blue dots indicating the same position on the slides at different time points. Scale bar = 50 µm