Figure 5.
14-3-3 θ binds to Cx43 phosphorylated in vivo. (a) His-14-3-3 θ bound to the GST-Cx43CT fusion protein and to Cx43 phosphorylated endogenously in Rat-1 cells as demonstrated here in an overlay assay. Proteins were resolved on a 12% polyacrylamide SDS-PAGE gel and transferred to an Immobilon-P membrane that was probed with His-14-3-3 θ and developed with an anti-His tag antibody. (b) GST-14-3-3 θ, but not GST alone, bound to Cx43 in cell lysates from EGF-treated and untreated Rat-1 cells as shown here. The GST input proteins are shown in the upper blot and the pull-down of Cx43 in the lower blot.