Fig. 5. NF-kB activation in TNF-α treated monocytes was inhibited by triacsin C or ACSL1 deficiency. Monocytic cells were pretreated with ACSL1 inhibitor (Triacsin C: (1uM) or vehicle for 1hr and then incubated with TNF-α for 15 minutes. NF-kB phosphorylation was determined by (A) western blotting and (B) flow cytometry. (C) Representative flow cytometry dot plots of p-NF-kB fluorescence versus total IkBα cells. (D) NF-kB reporter monocytic cells were pretreated with ACSL1 inhibitor (triacsin C: (1uM) or vehicle for 1 hour and then incubated with TNF-α for 24 hours. Cell culture media were assayed for SEAP reporter activity (degree of NF-κB activation). (E) ACSL-1 deficient cells were treated with TNF-α for 15 minutes. NF-kB phosphorylation was determined by (E) western blotting and (F) flow cytometry. (G) Representative flow cytometry dot plots of p-NF-kB fluorescence versus total IkB α cells. All data are expressed as mean ± SEM (n ≥ 3). *p<0.05, **p<0.01, ***p<0.001 versus TNF-α.