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Fig. 1. PKB binds periplakin. (A) The yeast two-hybrid fragments of PKB (fish) and periplakin (bait). Periplakin consists of six {alpha}-helical subdomains (aa 16-920), a rod domain (aa 945-1554) and a linker domain (L; aa 1556-1756). (B) GST pull-down assay with GST-c-ppl. A14 cells were transiently transfected with either wild-type or kinase-dead HA-tagged PKB. Lysates were incubated for 1 hour with GST-c-ppl. Samples were analysed for the presence of PKB by western blotting using the anti-HA antibody. A direct immunoprecipitation of HA-PKB using anti-HA was used as a control. The experiment was also performed with either GST or GST-c-ppl (right panel). (C) COS-7 cells were transiently co-transfected with myc-tagged c-ppl and either HA-tagged PKB or p70S6K. Co-immunoprecipitations were performed with anti-HA and anti-myc antibodies and immunocomplexes were resolved with SDS-PAGE and immunoblotted with anti-myc and anti-HA antibodies, respectively.