Extended Data Fig. 7
Lgr5pos cells are not required for the maintenance of 2D culture(a) Representative FACS gating of wildtype 2D and organoid cultures used to quantify Lgr5pos cells in Fig. 5a. (b) Joint t-SNE embedding of isogenic 2D and organoid cells showing expression of the isthmus marker Birc5, the pit marker Tff1, and the base/neck marker Aqp5. (c) Quantification of proliferation by FACS CellTrace dye detection following ablation of Lgr5pos cells in 2D culture (top) (n = 3 independent experiments, representative quantification shown). Representative phase contrast images of Lgr5DTR-GFP 2D cultures + /- DT continually for 10 passages, scale bars, 50 μm (n = 3 independent experiments). (d) Lgr5DTR-GFP organoids were treated with DT for 5 passages or passaged using standard conditions. Number of visible organoids were counted manually and recorded before splitting 1:3. Horizontal lines indicate mean number of organoids for each passage, n = 8 independent experiments. Representative FACS plots and representative images of organoids quantified at p5 are shown below and on the right of the graph, respectively.
