Skip to main content
. 2018 Jul 4;7(1):1–17. doi: 10.1016/j.jcmgh.2018.06.008

Figure 1.

Figure 1

A focused screen of IBD-related cytokines shows that IL22 causes a dose-dependent decrease in organoid survival and an increase in organoid size. (A–D) Ileal organoid screen for cytokine effects on intestinal epithelium. (A) Schematic of experimental design in which ileal organoids were treated for 6 days with 100 pmol/L IL6, IL17, IL21, or IL22. (B) Representative images of treated organoids after 6 days. Scale bar: 100 μm. (C) Percentage change in area of organoids comparing day 0 with day 6. Technical replicate n = 10+ organoids; biological N = 3 mice; significance is relative to untreated control. (D) Organoid efficiency relative to control organoids. Technical replicate n = 3 wells; biological N = 3 mice; significance is relative to untreated control. (E) Organoid response to a range of concentrations of IL22 (0, 0.8, 4, 20, 100, and 500 pmol/L), measured by change in organoid area (left Y axis, black lines, technical replicate n = 10+ organoids) and organoid survival (right Y axis, red lines, technical replicate n = 3+ wells) after 6 days. Biological N = 3 mice/treatment; significance is in relation to 0 pmol/L IL22. (F) Organoid survival with 0, 60, or 500 pmol/L IL22 treatment. Technical replicate n = 3 wells; biological N = 3 mice. Asterisks denote significance between the treatment group and control at the designated time point. (G and H) Organoids were treated with 500 pmol/L IL22 in the presence of 206 ng/mL IL22 neutralizing antibody or 206 ng/mL IgG of the same species. (G) Top: Representative images of treated organoids. Bottom: Quantification of organoid area. Technical replicate n = 10+ organoids; biological N = 3 mice; significance is relative to IgG-only control. (H) Top: Representative Western blot images for pSTAT3 and β-actin. Scale bar: 100 μm. Bottom: Quantification of intensity of Western blot bands normalized within each blot to the band with the highest intensity. Technical replicate n = 3 blots, biological N = 3 mice. Significance was calculated by 1-way analysis of variance with a Bonferroni correction for multiple comparisons. *P < .05, **P < .01, ***P < .001, ****P < .0001. pSTAT3, phosphorylated signal transducer and activator of transcription 3.