Figure 8

Analysis of the pathway by which loss of CLDN4 activates EphA2. (A) Western blot analysis showing no changes of active ?-catenin levels when EphA2 was knocked down in the CLDN4KD cells. (B) Representative Western blot showing the level of pAkt(Ser473), pEphA2(Ser897) and dephospho-?-catenin in the CLDN4KD cells untreated or treated with 50 μM PI3K inhibitor LY294002 for 1 h. (C) The histogram showing the mean level of pAkt(Ser473), pEphA2(Ser897) and dephospho-?-catenin expressed as the ratio relative to that in untreated CLDN4KD cells after normalization for their respective total protein. Results presented are mean ± SEM (n = 3). ^***p?<?0.001, ^*p?<?0.05 versus untreated CLDN4KD. (D) Representative Western blot showing that active ?-catenin was increased in the CLDN4KD cells (top panel) and siRNA-mediated knockdown of ?-catenin reduced the levels of pAkt(Ser473) and pEphA2(Ser897) in the CLDN4KD cells (bottom panel). (E) Histogram showing the mean levels of pAkt(Ser473) and pEphA2(Ser897) determined from 3 independent experiments expressed as the fold change relative to that in the scrambled siRNA transfected CLDN4KD cells after normalization to ?-actin. Results are presented as mean ± SEM (n = 3). ^###p?<?0.01, ^##p?<?0.01 versus scrambled siRNA transfected CLDN4KD.