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Fig. 5. SRD phosphorylation by endogenous CK2 inhibits homeoproteins intercellular transfer. (A) Extracts of COS-7 cells expressing cEN2{Delta}(146-169)SRD alone were separated by 2D gel and immunoblotted with anti-Engrailed serum. The acidic spot characteristic of wild-type cEN2 hyperphosphorylation is found (arrowhead). The asterisk indicates the position of the most basic isoform of wild-type cEN2. (B-E) Intercellular transfer of cEN2{Delta}(146-169)SRD (B) or cEN2SRD (C), cEN2SRD/5A (D), cEN2SRD/5E (E) expressed alone was analysed as in Fig. 2. Intercellular transfer of is inhibited independently of kinase overexpression (B). Ectopic addition of cEN2 SRD at the C terminus of wild-type cEN2 has the same inhibitory effect on intercellular transfer (C). S->A substitutions within the ectopic SRD restores transfer (D) but S->E substitutions does not (E). Arrowheads indicate transfected COS-7 cells. Scale bar: 30 µm.