Fig. 6
From: Integrin activation by the lipid molecule 25-hydroxycholesterol induces a proinflammatory response
The 25HC-integrin-FAK signaling network is required for optimal Nod2 response. a IL-6 in serum of mice injected (via intraperitoneal route) with 25HC (50 mg/kg; 4 h) (n = 5). b, c, e, g–m Cells were treated with 25 µg/ml of MDP for 8 h and d, f mice were injected intraperitoneally with 20 mg/kg of MDP for 4 h. C25H expression, 25HC production, FAK activation, and proinflammatory response were assessed. b RT-PCR analyses of C25H expression in MDP-treated wild-type (WT) and C25H knockout (KO) BMDMs. c 25HC levels in the medium supernatant of MDP-treated BMDMs. d 25HC levels in the serum of WT mice treated with MDP (n = 4). e IL-6 secretion from WT and C25H KO BMDMs treated with MDP. f IL-6 levels in the serum of WT and C25H KO mice treated with MDP (n = 4). g Western blot and densitometric analyses of FAK activation (phospho-FAK, Tyr397) status in MDP-treated THP-1 macrophages. h IL-6 production from MDP-treated WT and FAK KO MEFs (mouse embryo fibroblasts). i IL-6 secretion from MDP-treated NR-9456 macrophages in the presence of either control IgG or α5β1 integrin blocking antibody (Ab). j RT-PCR analyses of TNF expression in MDP-treated THP-1 cells transfected with either control siRNA (con siRNA) or α5 integrin-specific siRNA (α5 siRNA). k TNF secretion from MDP-treated WT and β3 integrin-deficient (β3+/− cells) BMDMs. l TNF secretion from MDP-treated THP-1 macrophages in the presence of either control IgG or αvβ3 integrin blocking antibody. m Western blot and densitometric analyses of FAK activation (phospho-FAK, Tyr925) status in MDP-treated WT and C25H KO BMDM. RT-PCR images are representative from two independent experiments. The ELISA values (mean ± standard deviation) are representative from two or three independent experiments (n = 4). *p ≤ 0.05 using a Student’s t-test. The densitometric quantification values for phospho-FAK (p-FAK) immunoblot represent the ratio of phospho-FAK:actin and the fold-induction was calculated after normalizing with the control untreated (UT) group. The densitometric values represent the mean ± standard deviation from three independent studies. *p ≤ 0.05 using a Student’s t-test
