Fig. 7. Intracellular localization of ForC truncation mutants fused to GFP. (A) Full-length ForC and the truncated ForC mutants. Gray boxes in the full-length ForC indicate the FH3 and FH2 domains. Thick lines indicate the regions encoded by each mutant. All ForC constructs were tagged with GFP at their N-termini. Crown localization of each mutant in either fixed or live cells is indicated by `-' and `+' on the right. (B) Fluorescence micrographs of forC cells expressing the various GFP-ForC mutants. Cells were fixed and stained with rhodamine-phalloidin. The full-length protein (a) and the 1-633 (c), 1-468 (d) and 1-323 (e) mutants all localized at the crowns (indicated by arrows), whereas GFP-FH3 did not (b, the position of a crown is indicated by an arrowhead). (C) Fluorescence micrographs of living forC cells expressing the GFP-ForC-1-323 mutant (left) and GFP-ForC (right). Arrows indicate the crown localization of GFP-ForC-1-323, which includes the region from the first methionine of ForC to the end of the FH3 domain. Crown localization of full-length GFP-ForC was not detected without fixation.