Impaired recruitment of myeloid cells to sites of inflammation in a model of acute peritonitis. (A) Impaired accumulation of Gr-1/Mac-1 double-positive cells in the inflamed peritoneum of Fak^−/− mice. WT and FAK^−/− mice were given intraperitoneal injections of 4% thioglycollate. Peritoneal lavage was collected 4 hours after injection, and Gr-1 and Mac-1 expression was examined by flow cytometry. Dot blots represent Gr-1/Mac-1–positive cells in the peritoneal cavity of WT and Fak^−/− mice, analyzed following PBS or thioglycollate injection. (B) Quantitative analysis of the percentage of Gr-1/Mac-1 double-positive cells in the peritoneal cavity of WT and Fak^−/− mice after PBS or thioglycollate injections (n = 3; *P < .05). (C) Quantitative analysis of the total number of Gr-1/Mac-1–positive cells recruited into peritoneal cavity of WT and Fak^−/− mice after PBS or thioglycollate injections (n = 3; *P < .05). (D) DNA was extracted from peritoneal cavity-derived cells and analyzed by PCR. Cre-mediated deletion of Fak was detected as a 327-bp fragment, and WT Fak was observed as a1.6-kb fragment. Lanes 1, 2, and 3 represent WT Fak bands after 4 hours of thioglycollate treatment in peritoneal cavity-derived cells. Lanes 4, 5, and 6 shows Fak deletion in peritoneal cavity-derived cells after 4 hours of thioglycollate treatment. (E) WT and FAK^−/− mice were given intraperitoneal injections of 4% thioglycollate. Peritoneal lavage was collected 4 days after injection, and F4/80-positive cells were detected by flow cytometry. Upper left quadrant of each dot blot represents the percentage of phycoerythrin-conjugated F4/80-positive cells in WT and FAK^−/− peritoneum after PBS or thioglycollate injections. (F) Quantitative analysis of the percentage of F4/80 positive cells in the peritoneal cavity of WT and Fak^−/− mice after PBS or thioglycollate injections (n = 3; *P < .05, 4 pairs of WT and Fak^−/− mice). (G) Quantitative analysis of the total number of F4/80 cells recruited to the peritoneal cavity of WT and Fak^−/− mice after PBS or thioglycollate injection (n = 3; *P < .05, 4 pairs of WT and Fak^−/−). (H) DNA was extracted from peritoneal cavity-derived cells and analyzed by PCR. Cre-mediated deletion of FAK was detected as a 327-bp fragment, and the WT Fak allele was observed as a 1.6-kb fragment. Lanes 1, 2, and 3 represent WT Fak bands in the peritoneal cavity-derived cells after 4 days of thioglycollate treatment. Lanes 4, 5, and 6 represent FAK deletion in cells recruited into the peritoneal cavity after 4 days of thioglycollate treatment.