KLF4 is essential for cell polarity and crypt-cyst formation in 3D culture of Caco-2 cells.

<p>(A) Immunofluorescent staining of Caco-2 cells in 2D and 3D culture with anti-α6-integrin and ZO-1 antibodies. (B) Caco-2 cells in 3D culture were stained for differentiation markers indicating cell polarity and cyst formation (definition of lumen-cyst vesus cyst structures). (C) Left: western blotting showing knockdown of KLF4 in Caco-2 cells. Right: statistical analysis of percentage of lumen-cyst formation in Caco-2 cell 3D cultures, comparing between control and KLF4 siRNA-transfected cell cultures. (*, P<0.05) (D) Left: western blotting showing expression of KLF4 in 293T cells co-transfected with human KLF4 and KLF4 shRNA plasmids. Right: statistical analysis of percentage of lumen-cyst formation in Caco-2 cell 3D cultures, comparing between control and KLF4 shRNA-infected cell cultures. (*, P<0.05).