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. Author manuscript; available in PMC: 2010 Aug 10.
Published in final edited form as: Nat Med. 2009 Apr 27;15(6):701–706. doi: 10.1038/nm.1951

Figure 5.

Figure 5

Putative intestinal stem cell populations with or without R-spondin1 treatment in culture. (a,b) Jejunal cultures contain Bmi1 lineage–derived cells. (a) Whole-mount LacZ staining of intestinal spheres. Bmi1+ cells and their progeny are identified as blue LacZ+ patches in RSpo1-Fc treatment versus vehicle control. (b) Frozen section of LacZ-stained intestinal spheres showing LacZ+ Bmi1 lineage–derived cells and their progeny in the epithelial layer of RSpo1-Fc-treated cultures. (c) Fluorescent in situ hybridization (FISH) for Lgr5 (orange) with simultaneous DAPI nuclear stain (blue) at culture day 35. (d) Percentage of Lgr5+ cells as a fraction of epithelial cells after RSpo1-Fc treatment compared to vehicle control at day 35. RSpo1-Fc was added once weekly for 5 weeks. Error bars indicate s.e.m.; n ≥ 4. *P < 0.05 versus vehicle control. (e) In situ hybridization (ISH) for Lgr5 in vivo from mice 7 d after single intravenous administration of adenoviruses encoding RSpo1-Fc (Ad RSpo1-Fc) or antibody Fc control (Ad Fc). (f) Histology of jejunum and colon from Ad RSpo1-Fc–treated– or Ad Fc–treated mice. H&E and PCNA staining of intestine 7 d after single intravenous injection of Ad RSpo1-Fc or Ad Fc is depicted (e,f). Scale bars, 100 μm.