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Figure S3. Large, proliferative, duct-derived clones as opposed to small, abortive, lobule-derived clones are multipotent. (A) Three-color flow cytometric detection of SSEA-4, EpCAM, and CD49f in uncultured Lin^- epithelial cells. (a) Dot plot shows SSEA-4^hi cells (black dots) within a Lin^-/CD49f/EpCAM context (green dots). (b) Dot plots of SSEA-4 against CD49f to illustrate the magnitude of difference in SSEA-4 expression between subpopulations I, II, and III + IV. SSEA-4^hi cells clearly enrich within the CD49f⁺/EpCAM^hi (II) gate. The percentages of SSEA-4^hi cells within each subpopulation are indicated in the rectangular gates. (B) Multicolor imaging of a clone derived from a single FACS cloned SSEA-4^hi cell stained with keratin K19 (red) and keratin K14 (green). Bar, 50 µm. (C) Isolation of the four subpopulations using EpCAM directly conjugated to FITC. Application of EpCAM directly conjugated to FITC reveals a FACS profile comparable with the profile obtained when using unconjugated antibodies visualized with fluorescent secondary antibodies.