By staining the spheroids with p68, FITC-phalloidin for F-actin and DAPI (for nuclear staining) to visualize spheroid architectures under an inverted confocal microscopy, we found that cyst lumens which were surrounded with polarized epithelial cells as outlined by F-actin to the mark the membrane were always formed on mIMCD3 cell (control siRNA) 3D cultures, whereas the lumens failed to be expanded as seen with an undetectable or markedly diminished luminal space in the 3D cultures with p68 knockdown cells (Figure 8C). Here, DDX5 is linked to cyst.