Fig. 6. Requirement of Par-6 for co-localization of afadin with nectin. (A) Reduction in the amount of Par-6 by RNAi. The lysates of Par-6-knockdown MDCK cells were subjected to SDS-PAGE, followed by western blotting with the indicated Abs. (B) Reduction in the immunofluorescence signals for E-cadherin, afadin, claudin-1, occludin, JAM-A, Par-3 and ZO-1 at cell-cell adhesion sites in Par-6-knockdown MDCK cells. After Ca²⁺ switch, the cells were fixed and stained with various combinations of the indicated Abs. (C) Reduction in the immunofluorescence signal for afadin, but not that for nectin-1, at the nectin-based cell-cell adhesion sites in Par-6-knockdown nectin-1-MDCK cells. After Ca²⁺ switch, the cells were fixed and stained with various combinations of the indicated Abs. Bars, 10 µm.