Fig. 5. PIX and ß1PIX induced cell ruffling via Rac1. (A) PIX and ß1PIX promote the formation of phase-dark ruffle-like structures. HeLa cells were microinjected with DNA constructs of the various PIX and PIX mutants in pXJ-40 vector. The cells were then viewed by phase-contrast microscopy. The injected cells are marked by arrowheads. The DNA construct used were pXJ-Flag-PIX, pXJ-Flagß1PIX, pXJ-Flag-ß2PIX, pXJ-Flag-N80ß1PIX, pXJ-Flag-ß1PIX^1-555, pXJ-GST-ß1PIX-C-ter, pXJ-Flag-ß1PIX-DHm and pXJ-GST vector. (B) The formation of these ruffles could be blocked by Rac1^N17 but not Cdc42^N17. HeLa cells were microinjected with pXJ-40 DNA construct of ß1PIX together with Rac1^N17 or Cdc42^N17. The injected cells are marked by arrow heads. (C) ß1PIX and ß2PIX could induce filopodia and cell rounding when wild-type Cdc42 was also overexpressed. HeLa cells were microinjected with the plasmid DNA constructs in pXJ-40 vector and the cells were observed 2-2.5 hours after microinjection. The plasmid DNA injected are: 1, hPem2; 2, Cdc42^V12; 3, ß1PIX and Cdc42 wild type (w/t); 4, ß2PIX and Cdc42 w/t; 5, Cdc42 w/t alone.

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