a, Cystic spheroids formation by CRAD KO. Isolated crypts from WT, CRAD KO, and APCMIN were maintained in the organoid culture medium. These data are representative of three independent organoid experiments with similar results. 10 organoids per group [WT vs. KO] were analyzed.
b-l, IHC analysis of the organoids derived from CRAD WT and KO mouse intestine. Compared to WT, CRAD KO-driven cystic spheroids showed that increase of cell proliferation (Ki67; b), increase of β-catenin (c) and its target genes (Cyclin D1 [d]; CD44 [e]; MYC [f]), disruption of the actin cytoskeleton (Phalloidin [g]; Actin [h]), loss of epithelial cell integrity (CK19 [i]; Villin [j]), and decreased IEC lineage differentiation (Chromogranin A: ChgA; [k]; Lysozyme [l]).
Representative images of three experiments; Red scale bars indicate 20μm; White scale bars indicate 20μm.