Figure 6.

CD44⁺CD24^loCD166⁺ GRP78^−/lo identifies ISCs independent of Lgr5-GFP^hi cells. (A) The mosaic distribution of Lgr5-GFP⁺ crypt in jejunum. Gradient GFP expression in Lgr5-GFP⁺ crypt. (B) Lgr5-GFP^neg, Lgr5-GFP^lo/med, and Lgr5-GFP^hi gates in the CD44⁺CD24^loCD166⁺ GRP78^−/lo population. (C) qRT-PCR results show the gene expression of Lgr5-GFP^neg and Lgr5-GFP^hi cells relative to Lgr5-GFP^lo/med cells from the CD44⁺CD24^loCD166⁺ GRP78^−/lo population. Data are presented as mean ± SEM (n = 4, *P < .05, **P < .01, ***P < .001). (D) CFE analysis of the Lgr5-GFP^neg, Lgr5-GFP^lo/med, and Lgr5-GFP^hi cells in the gates of B. Data are represented as mean ± SEM (*P < .05, n = 3, t test). (E) A growth process of single cells sorted from Lgr5-GFP^neg, Lgr5-GFP^lo/med, and Lgr5-GFP^hi gates in B, showing that Lgr5-GFP^lo/med cells can generate Lgr5-GFP^hi enteroid. (F) FACS analysis of enteroids derived from single Lgr5-GFP^neg, Lgr5-GFP^lo/med, and Lgr5-GFP^hi cells, showing that single Lgr5-GFP^lo/med cells can generate enteroids containing Lgr5-GFP^hi cells as single Lgr5-GFP^hi cells did. Data are presented as mean ± SEM (n = 3, **P < .01).