Figure 2
Bulk RNA-sequencing analysis of organoids stimulated with norepinephrine or UK 14,304. (A) Experimental set-up. Mouse small intestinal organoids were grown and after four passages they were stimulated with either norepinephrine or UK 14,304 1 μM (or vehicle controls) for four days. Afterwards they were harvested and processed (n = 5). (B,C) Heat map visualizing the top 30 upregulated and downregulated genes upon stimulation with norepinephrine (B) or UK 14,304 (C) and vehicle control. (D) Venn diagrams showing overlap of upregulated and downregulated genes upon stimulation with norepinephrine or UK 14,304. (E) Single gene expression analysis of intestinal cell markers Bmi1 (P = 0.089), Olfm4 (P = 0.116), Lyz1 (P < 0.0001), Muc2 (P = 0.927), and Tff3 (P = 0.160) in norepinephrine versus vehicle control treated organoids. (F) Single gene expression analysis of intestinal cell markers Bmi1 (P = 0.217), Olfm4 (P < 0.0001), Lyz1 (P < 0.0001), Muc2 (P = 0.089), and Tff3 (P < 0.0001) in UK 14,304 versus vehicle control treated organoids. (G,H) Single gene expression analysis of cell proliferation marker Mki67 in norepinephrine (G; P = 0.100) or UK 14,304 (H; P = 0.0001) versus vehicle control treated organoids. (I,J) Gene Set Enrichment Analysis (GSEA) of cell type signatures in norepinephrine (I) or UK 14,304 (J) treated organoids versus vehicle controls. Blue bars indicate adjusted P < 0.05, and red bars indicate adjusted P > 0.05. (K) GSEA of signature of stem cells in UK 14,304 treated organoids versus vehicle controls (Normalized enrichment score (NES) = 1.48; P = 0.006). (L) GSEA of signature of Paneth cells in UK 14,304 treated organoids versus vehicle controls (NES = − 2.61; P = 0.002). (M) GSEA of signature of goblet cells in UK 14,304 treated organoids versus vehicle controls (NES = − 2.22; P = 0.003). Statistical significance is indicated as follows: *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. Data are shown in boxplots.
