Extended Data Fig. 6: Flow cytometry analysis strategy of lineage-traced organoids. | Nature Cell Biology

Extended Data Fig. 6: Flow cytometry analysis strategy of lineage-traced organoids.

From: Chromatin remodelling in damaged intestinal crypts orchestrates redundant TGFβ and Hippo signalling to drive regeneration

Extended Data Fig. 6

a) Schematic of experimental design accompanying Fig. 4d. Clu-CreERT2;lsl-tdTom;Lgr5-GFP organoids were treated with a pulse of TGFβ (TGFβ1; 24 h) and 4-OHT to induce recombination prior to passaging and flow cytometry. b) Gating strategy used to quantify cell populations isolated from organoids via flow cytometry. Plots indicate cell populations that were included based on size (singlets) and viability (DAPI-). c) Representative plots indicating the proportion of (1) GFP-/tdTom- (2) GFP + /tdTom-; (3) GFP + /tdTom+ and (4) GFP-/tdTom+ populations in Clu-CreERT2;lsl-tdTom;Lgr5-GFP organoids treated +/-4-OHT to induce lineage tracing and +/-TGFβ1 (1, 50 or 200 pM), as indicated (bottom panels). GFP-/tdTom- and GFP-/tdTom+ organoids were used for gating controls.

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