Figure 6.

Igf2 Knockout Impairs the Growth of Intestinal Organoids

WT and iKO (Igf2^fl/flCre^+/−) organoids were cultured in medium containing EGF, Noggin, and R-spondin1 (ENR) for 5 days. Arrows delineate small organoids with few Paneth cells. Arrowhead indicates dying organoid on day 3, which was confirmed to be dead on day 5 (A). Survival curve of organoids from WT and iKO mice (B). 4-OHT or vehicle was added to iKO (Igf2^fl/flCre^+/+) organoid cultures at day 7. The organoids were imaged daily (C), harvested for genotyping (D), and scored daily (E). The positive control (p.c.) used in (D) is the genomic DNA sample from a Rosa26-CreER heterozygous mouse sample to show the WT (bottom) and mutant (upper) PCR products. EdU staining was performed at day 4 after 4-OHT treatment (F and G). Scale bars, 200 μm. ^∗p < 0.05, ^∗∗p < 0.01, ^∗∗∗p < 0.001 versus controls by Student's t test or one-way ANOVA. Data were analyzed from n = 3 independent mice. Error bars represent SEM.