Figure 1.
Suppression of Scrib expression in MDCK cells causes a delay in tight junction assembly. (A) MDCK II cells were transiently transfected with either pS-Luciferase vector (Luc) as a control or three pSUPER constructs targeting different sequences of canine Scrib mRNA (ScrbKD). Equal amounts of proteins were analyzed by SDS-PAGE and immunoblot. (B) Control and ScrbKD MDCK cells grown at high density were fixed and stained for Scrib and occludin. (C) Control and ScrbKD cells were subjected to a calcium switch and then fixed and stained for ZO-1 at the indicated times after readdition of calcium. Magnified details of the junctions are shown in inverted black-and-white scale to highlight the defects caused by loss of Scrib.