Fig. 2

WNT4 promotes symmetric fission of crypt in organoids. A, B Representative time-lapse images of mouse small intestinal organoids transfected with si-control, si-Wnt4, Vector or Wnt4-OE, or treated with an inhibitory Eph fragment (Eph, 10 μg/ml) at the indicated time points. Scale bars, 100 μm. Red dashed boxes indicate symmetric fissions, and yellow dashed boxes indicate asymmetric fissions. C, D Representative 3D projections of organoids stained against F-actin (Phalloidin, red) and nuclei (Hoechst, blue) after transfection with si-control, si-Wnt4, Vector or Wnt4-OE, or treated with an inhibitory Eph fragment (Eph, 10 μg/ml). Scale bars, 50 μm. E, F The symmetry ratio was calculated by dividing the length of the short daughter crypt by the length of the long daughter crypt based on the 3D projections of organoids in C and D. n = 24 for si-control, n = 29 for si-Wnt4, n = 29 for Eph, n = 27 for vector, and n = 20 for Wnt4-OE. All values are means ± SD. Statistical analyses were performed by one-way ANOVA with Tukey’s multiple comparisons test (E) or unpaired Student’s t-test (F). **p < 0.01, ****p < 0.0001