Figure 5.

Adaptive shift of stem cell phenotype under selective pressure

(A) Shift in stem cell-marker expression detected by qRT-PCR following exposure of wild-type organoids to increasing concentrations of media IFN-γ. Statistical analysis, t test, p values as stated.

(B) Skewed expression of stem cell markers, detected by FACS for Ly6a and GFP, following exposure of Lgr5-GFP labeled murine organoids to 5 μL/mL of media IFN-γ.

(C) Schematic showing timing of recombination, Diphtheria Toxin (DT) activation, and tissue harvesting of Lgr5^DTR;Apc^Min mice.

(D) Using stem cell index to map polyp tissue from Apc^Min and Lgr5^DTR;Apc^Min to show dynamic change in stem cell molecular phenotype measured by stem cell index, before and after DTR activation and CBC cell ablation.

(E) Gene set enrichment analysis showing enrichment of Ifn-γ signaling and Yap signaling between day 0 (unrecombined) and day 1 (after DTR stem cell ablation).

(F) Dual-color ISH for Lgr5 (CBC marker, green) and Anxa1 (RSC marker, red) to show marker expression change in Apc^Min and Lgr5^DTR;Apc^Min polyps before and after CBC ablation. Scale bars, 100 μm.