Fig. 6

Long-term competitive reconstitution of HSCs based on prospective classification by the LSM model. A LSK/Evi1-GFP⁺ cells were FACS-sorted from Evi1^GFP transgenic mice and classified by the LSM model. A representative field of view is shown. Scale bar = 10 μm. B GFP fluorescence analysis of the classified cell types. Solid green dots are individual GFP⁺ cells and the gray outlines depict the distribution of their GFP fluorescence intensity. The orange dashed lines indicate the medians of each cell groups. Increasing the prediction score threshold from 0.34 (all scores) to 0.9 didn’t significantly change fluorescence intensity features among predicted cell types, and the cells with the highest GFP expression (inside red boxes) were always classified as LT-HSC. n = 5. C Competitive reconstitution of irradiated mice with GFP-high cells. Top 3% high GFP expressing cells were FACS-sorted from the LSK/Evi1-GFP⁺ pool and transplanted into lethally irradiated recipients. Each recipient was transplanted with 5 or 10 GFP-high cells and 3 × 10⁵ host-derived BM cells. GFP-negative LSK cells were used as control. BM was harvested after 4 months for chimerism analysis. n = 3 each group. **p < 0.01 (compared to GFP-negative group. Unpaired t test)