To validate the stem cell status of the Lgr5-expressing CBC cells, a transgenic mouse model was generated with an eGFP reporter gene and a gene encoding a tamoxifen-regulated Cre protein inserted downstream of
We then labeled Lgr5+ cells with the Rosa26-LacZ reporter in the Lgr5EGFP-Ires-CreERT2; Rosa26-LacZ mice, and further determined if new retinal cells could be generated from labeled cell at later times. Results:We observed that Lgr5 was expressed in the inner nuclear layer of the ...
a,Lgr5-EGFP-IRES-creERT2knock-in mouse crossed withRosa26-lacZreporter mice 12 h after tamoxifen injection.b, Frequency at which the blue cells appeared at specific positions relative to the crypt bottom, according to the scheme in the inset. Results are depicted as means and standard devi...
Inhibition of BMP signaling in Lgr5 cells was achieved by crossing Lgr5-EGFP-ires-CreERT2 (Lgr5-Cre) mice to mice with floxed alleles of BMP receptor 1A (Lgr5-Cre;Bmpr1aflox/flox mice). Lgr5/GFP+ve cells were isolated using flow cytometry. Lineage tracing studies were conducted by ...
afterBmpr1ainactivation (Supplementary Fig. 1g). To examine whether loss of BMP response in Lgr5+ISCs is responsible for their expansion, we deletedBmpr1aspecifically in these cells usingLgr5-EGFP-IRES-CreERT2;Bmpr1afl/fl(Lgr5-CreER;Bmpr1afl/fl) mice. Indeed, ISC-specificBmpr1adeletion also...
We cultured crypts from Lgr5–EGFP–ires–CreERT2 mice crossed with the Cre-activatable Rosa26–LacZreporter to allowlineage tracing. Directly after induction with low-dose tamoxifen, we noted single labelled cells (Supplementary Fig. 4e, g). More than 90% of these ...
(E) Whole-mount intestine from E-cadherin-mCFP (white)/Lgr5-EGFP-Ires-CreERT2 (green) mice. Lgr5-GFPhi population was visualized in red (false color), whereas E-cadherin-mCFP (white) marks cell borders. At the crypt base, all Lgr5+ cells were GFPhi. (F) Counting in 3D reconstru...
eGFP-IRES-CreERT2 knock in allele, with mice carrying conditional alleles ofTrp53andRb1(akap53/TP53andRb/RB1), respectively. To trace cells exposed to tamoxifen-activated Cre-ERT2 fusion protein mice were also crossed toRosa-loxP-stop-loxP-tdTomato(Ai9) reporter mice (Supplementary Fig.2a)...
To investigate the functional role of Lgr5 and Lgr6 in adult lungs, we characterized Lgr6 expression in the lung using Lgr6-EGFP-IRES-CreERT2 knockin mice, in which EGFP marks cells with active expression of the Lgr6 locus (Snippert et al., 2010). Unexpectedly, rather than marking epithelia...
Genetically modified mice, which include Lgr5-EGFP-IRES-CreERT2 (Lgr5CreERT2/+) knock-in mice (Barker et al., 2007) and Rosa26-floxed stop cassette-tdTomato (R26RtdTomato) reporter mice were purchased from the Jackson Laboratory. The embryos of these mice were further investigated, as ...