K-Ras V12 and B-Raf V600E disrupt normal morphogenesis in Caco-2 3D cultures. (A) 3D cultures expressing control (pQCXIP GFP), K-Ras V12 (pQCXIP GFP K-Ras V12), B-Raf V600E (pQCFLAP B-Raf V600E), or PI3KCA H1047R (pQCFLAP PI3KCA H1047R) were fixed at day 10 and stained for aPKC and DNA. Representative images are shown. Bar, 50 µm. (B) The midplanes of ≥50 structures were imaged for each condition in three independent experiments and categorized as having normal, multiple layer, or solid morphology. The number of nuclei in the midplane of each structure was also quantified. The means ± SEM are shown. **, P < 0.01; ***, P < 0.0005; ****, P < 0.0001.