RationaleACSL catalyzes the ligation of fatty acid to coenzyme A, the first and rate limiting step for fatty acid utilization. Triacsin C is a fungal metabolite which inhibits ACSL1, ACSL3, and ACSL4. Earlier, we showed that Triacsin C reduces the infarct volume and inhibits iNOS expression in the MCAO model of stroke. As iNOS is up‐regulated in stroke, we wished to determine whether the Triacsin C effect was linked to iNOS expression.MethodsbEnd.3 cells were stimulated with cytokine mix, (TNFα, 60 ng/mL; Ilβ,2 ng/mL; IFNγ, 100 units/mL), in the presence or absence of 5μM Triacsin C for 24 hours. RNA was extracted following RNeasy Plus Mini Kit(Qiagaen) instruction. The concentration and purity of RNA was determined by Spectrophometer. 20 μl first strand cDNA was synthesized from 1 μg total RNA for each sample using Superscript III First‐Strand(Invitrogen). One μl cDNA was amplified using SYBR Green PCR Master Mix (applied Biosystems) in CFX96 Touch Real‐Time PCR Detection System (BioRad). β‐actin (cat 330001 PPM02945B) and iNOS (cat 330001PPM02928B) primers were purchased from Qiagen. Expression level in fold change was determined by the comparative threshold cycle method (2−ΔΔCt) with β‐actin as the control gene.ResultsCytomix stimulated iNOS expression in the presence of Triacsin C was 52.8 ± 8.33% of control. (p=0.0005; n=5 independent experiments).ConclusionsOur data confirm that iNOS expression is modulated by ACSL activity, and support our hypothesis that the Triacsin C effect on stroke infarct volume is related to suppression of iNOS expression.Support or Funding InformationDepartment of Pharmaceutical Sciences, Texas Tech University Health Sciences Center, Amarillo, TXThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.