Morphological and biochemical analysis of Rac1 in three-dimensional epithelial cell cultures
Journal
Methods in Enzymology
Journal Volume
406
Pages
676-691
Date Issued
2006
Author(s)
Abstract
Rho GTPases are critical regulators of epithelial morphogenesis. A powerful means to investigate their function is three-dimensional (3D) cell culture, which mimics the architecture of epithelia in vivo. However, the nature of 3D culture requires specialized techniques for morphological and biochemical analyses. Here, we describe protocols for 3D culture studies with Madin-Darby Canine Kidney (MDCK) epithelial cells: establishing cultures, immunostaining, and expressing, detecting, and assaying Rho proteins. These protocols enable the regulation of epithelial morphogenesis to be explored at a detailed molecular level.
Publisher
Academic Press Inc.
Type
review