FIG. 4.

Overexpression of FRNK, Phe397 FAK, and Arg454 FAK inhibited the E. coli invasion of HBMEC. Cell lysates of HBMEC that had been transfected with plasmids containing GFP (Control or C) or HA-tagged FRNK (A) and FAK-WT, Phe925, FAK, Phe327 FAK, or Arg454 FAK (B) were immunoprecipitated with anti-HA tag antibody. The immune complexes were subjected to immunoblotting with anti-FAK antibody. The arrow indicates the FRNK protein (A). The protein above the FRNK band was due to immunoglobulin G present in the immune complexes. (C) HBMEC transfected with FAK mutants were used for invasion assays as described in Materials and Methods. Values are means ± standard deviations of results from at least five separate experiments carried out in triplicate, and invasion is expressed as a percentage of the value for GFP-expressing cells (control). P < 0.01 for FRNK, Phe397 FAK, and Arg454 FAK for E44 compared to control cells by unpaired two-tailed t test.