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Figure 4


Fig. 4. Effects of PKA inhibitors on PCLN-1 distribution. Mock and FLAG-tagged PCLN-1-expressing MDCK cells were treated with 50 µM H-89, 10 µM PKI, 50 µM DDA or 50 µM DDA for 1 hour. When used, 100 µM 8-Br-cAMP (cAMP) was treated for 1 hour after the addition of DDA. (A) The Triton X-100-soluble (60 µg) and -insoluble fractions (30 µg) were immunoblotted with anti-FLAG antibody. (B) The whole membrane fractions (500 µg) were incubated with protein G-Sepharose and anti-FLAG antibody to immunoprecipitate (IP). The immune pellets were immunoblotted (IB) with anti-ZO-1 antibody (upper) or anti-PCLN-1 (lower) antibodies. Whole membrane fractions (30 µg) were immunoblotted with anti-ZO-1 antibody (middle).





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