Figure 8.

Early progeny cells reacquire a stem cell–like transcriptional profile after DXR. (A) Experimental design to assess transcriptional differences in early progeny vs aISCs in control Lgr5^eGFP-CreERT2 reporter mice. Control mice did not receive any additional treatment. Crypt-enriched epithelium was sorted for microfluidic PCR analysis by FACS on the basis of GFP and tdTomato fluorescence (GFP⁺, tdTomato⁺, GFP⁺/tdTomato⁺, negative). (B) Mean log2 fold change of aISC and reserve intestinal stem cell (rISC)–associated transcripts in the indicated populations isolated from Lgr5^eGFP-CreERT2 reporter control mice 24 hours after TAM injection (n = 3, normalized to double negative epithelium). (C) Experimental design to assess early transcriptional responses to DXR treatment in early progeny vs aISCs. Mice were injected with DXR 1 day after TAM injection and FACS was performed on crypt-enriched epithelium at 6 or 24 hours post-DXR, similar to Figure 6A. (D) Mean log2 fold change of aISC or rISC-associated transcripts in the indicated populations 6 hours (n = 2) and 24 hours (n = 3) after DXR. Each cell population was normalized to its control population (Figure 6B) to demonstrate DXR-specific responses within the identified cell type. Statistically different gene expression (P values in text) was calculated by 2-way analysis of variance and the false discovery rate was controlled with the 2-stage step-up method of Benjamini, Krieger, and Yekutieli.