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Fig. 7. Requirement of aPKC for the association of afadin with nectin. (A) Reduction in the amount of aPKC by RNAi. The lysates of aPKC-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 aPKC-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 aPKC-knockdown nectin-1-MDCK cells. After Ca2+ switch, the cells were fixed and stained with various combinations of the indicated Abs. Bars, 10 µm.