In the confocal images, the GFP fusion proteins are represented by green, the TR-WGA is represented by red and the Hoechst nuclear stain is represented by blue. TR-WGA and Hoechst stain. Three distribution patterns are generated from the GFP fusion proteins. The GFP fusion proteins are represented by green, the TR-WGA is represented by red and the Hoechst stain is represented by blue. Open arrowheads point to the outer segment plasma membrane while filled arrowheads point to the inner segment plasma membrane.
Membrane association is required for outer segment transport.
For our C-terminal fusion proteins, this was achieved via palmitoylation. Deletion of the terminal 5 amino acids or mutation of the penultimate proline results in delocalization of the fusion protein. This contrasts with results in MDCK cells by Chuang et al. This difference may reflect the differences in cell type used as the expression system. We will determine whether any of the distribution patterns typified by Fig. 1 represent default transport pathways.