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. 2012 Feb 28;31(8):1999–2012. doi: 10.1038/emboj.2012.49

Figure 3.

Figure 3

CLDN14 interacts with CLDN16. (A) Y2H assays showing interaction of CLDN14 with CLDN16 but not with CLDN19, determined by three reporter genes (HIS3, lacZ, and ADE2) in the yeast NMY51 strain. Plates with selective medium lacking leucine and tryptophan (SD-LW), indicating the transformation of both bait and prey vectors; with SD-LWH and SD-LWHA, indicating the expression of reporter genes HIS3 and ADE2; and the β-galactosidase assay are shown. (B) Quantitative β-galactosidase assay to determine the relative interaction strength of the following pairs: CLDN16–CLDN19, CLDN14–CLDN16, and CLDN14–CLDN19. (C, D) Coimmunoprecipitation of claudins in doubly transfected HEK293 cells to determine CLDN14–CLDN16 interaction (C) and CLDN14–CLDN19 interaction (D). (E) Coimmunoprecipitation of claudins in triply transfected HEK293 cells to determine CLDN14–CLDN16–CLDN19 interaction. Antibodies used for immunoprecipitation are shown above the lanes; antibody for blot visualization is shown at left. Non-specific 1: anti-CLDN1; Non-specific 2: anti-occludin; *Lane shows 10% of input amount as in other lanes.