Architecture, expression profile and subcellular localization of GxcDD. (A) Schematic representation of the domain organization of GxcDD and the domain constructs used in the study. (B) Northern blot showing the expression profile of GxcDD during Dictyostelium development on phosphate agar plates. Cells were harvested at the indicated time points and RNA isolated by phenol-chloroform and probed using a fragment derived from the 3' end of the GxcDD cDNA (nt 3807–4860). (C) Western blot showing the accumulation of GxcDD during development. Total cellular proteins were harvested at the indicated time points and subjected to western blot analysis using polyclonal antibodies raised against GxcDD. (D) (top) Membrane (P) or cytosolic fractions (S) and (bottom) Triton X-100 soluble (S) and insoluble (P) cytoskeletal fractions were prepared as described in Materials and methods and subjected to immunodetection using GxcDD polyclonal antibodies.