Fig. 5
From: Integrin activation by the lipid molecule 25-hydroxycholesterol induces a proinflammatory response
α5β1 and αvβ3 integrins regulate 25HC-mediated proinflammatory response. a, c–g α5β1 and αvβ3 integrin-deficient cells were treated with 50 µM 25HC for 8 h to evaluate NFκB activation and proinflammatory response. a TNF secretion from 25HC-treated NR-9456 macrophages in the presence of either control IgG or α5β1 integrin blocking antibody (Ab). b Western blot analyses of α5 integrin protein expression in THP-1 cells transfected with either control siRNA (con siRNA) or α5 integrin-specific siRNA (α5 siRNA). c RT-PCR analyses of TNF expression in 25HC-treated THP-1 cells transfected with either control siRNA (con siRNA) or α5 integrin-specific siRNA (α5 siRNA). d Western blot and densitometric analyses of phospho-IκB status in 25HC-treated wild-type (WT) and α5 integrin knockout (KO) HAP1 cells. e TNF secretion from 25HC-treated WT and β3 integrin-deficient (β3+/− cells) BMDMs. f, g TNF (f) and IL-6 (g) secretion from 25HC-treated THP-1 macrophages in the presence of either control IgG or αvβ3 integrin blocking antibody (Ab). 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-IκB (p-IκB) immunoblot represent the ratio of phospho-IκB:actin and the fold-induction was calculated after normalizing with the control 0 min group. The densitometric values represent the mean ± standard deviation from three independent studies. *p ≤ 0.05 using a Student’s t-test
