Figure 3.

SH3BP1 regulates epithelial morphogenesis. (A–C) Caco-2 cells were transfected with siRNAs and then plated in a 3D matrix. After 5 d, the cells were fixed and first analyzed by phase-contrast microscopy (A) before staining with fluorescent phalloidin and Hoechst followed by confocal microscopy (B). Note that SH3BP1 depletion leads to smaller and often disorganized cysts. The effect of SH3BP1 depletion was quantified by counting normally organized cysts with a single lumen and disorganized cysts with no or multiple lumen (only cysts that had reached a minimal size of 30-µm diameter were considered for the morphological quantification to avoid a bias because of the smaller cysts). Shown are means ± 1 SD of three independent experiments (n = 3). Bars, 10 µm. p, phosphorylated.