TLR3 signaling regulates cell migration in a biphasic manner. A, Flag-TLR3-expressing HEK293 (Wt11) cells were treated with poly(I:C) (100 μg/ml) for the indicated times, cell lysates were immunoprecipitated with Flag antibody, and analyzed for pY416Src, Src, or TLR3 by Western Blot. B, HEK293 (293) or Wt11 cells were treated with poly(I:C) for 1 h when cell lysates were analyzed for pY416Src and actin by Western Blot. C, Wt11 cells were treated with poly(I:C) for 1 h, when cell lysates were analyzed for pY576/577FAK or FAK by Western Blot. D, Mouse bone marrow-derived macrophages (BMDMs) were treated with poly(I:C) for 1h, when the cell lysates were analyzed for pY416Src and pY576/577FAK by Western Blot. E, Mouse BMDMs were treated with poly(I:C) and their migration over 4 h or 24 h was measured by wound healing assay. For each time point, the number of cells that migrated in the untreated plates was considered as 100% and all other values were normalized to this. F, BMDMs were pre-treated with poly(I:C) for 4 h or left untreated, followed by wound healing assay with poly(I:C) treatment for another 4 h.