Fig. 2. Levels of ZO-1, ZO-2, claudin-6, occludin and Lfc proteins are altered following cingulin mutation in EBs. Triton-soluble and Triton-insoluble fractions of wild-type (+/+), heterozygous (+/–) and homozygous (–/–) EBs (genotype and clone name indicated above each lane) were analysed by SDS-PAGE and western blotting using antibodies against the proteins indicated on the left. Anti--tubulin signal was used to normalize protein loading. Numbers on the right indicate size (kDa) of protein markers. FL, full-length cingulin (140 kDa); T, truncated cingulin (100 kDa). Densitometric analysis was performed to measure protein levels when marked differences were observed. The numbers below the cingulin, ZO-1, ZO-2, claudin-6 and occludin lanes represent the signal intensity (mean of at least three independent experiments, one of which is shown here) relative to an arbitrary level of 1, corresponding to a reference lane for each set. ND, not detected.