Fig. 5.

FAK mediates injury-induced LIF and IL-6 induction in C6 cells. C6 cells were seeded and maintained for 48 h in serum-containing medium without added VTN. Cells were then injured in an in vitro trauma model (swipe injury) with or without FAK inhibitors added at the time of injury. LIF (A) and IL-6 (B) mRNA expression were strongly induced (Ctrl Inj) at 4 h after injury compared to no injury controls (Ctrl NI), but were abolished by treatment with FAK antagonists, PND-1186 (PND), PF573228 (PF228), PF562271 (PF271), but not Y11. Surprisingly, Y11 further increased IL-6 expression after injury. Data are means±s.e.m. of three independent experiments and expressed as a fold change relative to uninjured controls, first normalized to GAPDH to account for differences in cell numbers. *P<0.05; **P<0.01; ***P<0.001; NS, not significant. In the same conditions as described for A and B, LIF (C) and IL-6 (D) protein expression and release was increased after injury, and attenuated by all inhibitors, except Y11, as shown by dot blots of conditioned medium (representative of three independent experiments). In addition, we found that C6 cells overexpressing wild-type (WT) FAK expressed significantly more LIF (E) and IL-6 (F) than cells overexpressing dominant negative Y397F mutant FAK following injury (mean±s.e.m.; n=3).