Fig. 3.

DACH1 stimulated CRC organoid formation. a. Representative images of the CRC organoid spheres. Sphere areas decreased in all the DACH1-knockout organoids (transfected with 3 single guide RNAs, sgRNAs, designated as sg1, sg2 and sg3) compared with that in the NT group. The scale bars for the upper row=100 μm, and the scale bars for the lower row=200 μm. b. DACH1 was efficiently knocked out in sgRNA-transfected organoids, as detected by WB. c. Total sphere areas decreased in the DACH1 sgRNA-transfected group compared with those in the NT group. d-e. The organoid areas in the presence of DACH1 expression were within the lower quartile more than the control; scale bars =200 μm. f-g. The average sphere areas of DACH1 knockout organoids were decreased compared with those in the NT control; scale bar=100 μm. h-i. The sphere growth rates in NT organoids were higher than those in the DACH1-knockout group; scale bar=100 μm. j-k. The organoid forming efficiency was decreased by DACH1 knockout. The scale bars in the left column=100 μm, and the scale bars in the right column=200 μm. For 3c, 3 g, 3i and 3k, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, Student's t-test. Error bars: mean±SD.