FW1256 is a phenyl analogue and a slow-releasing hydrogen sulfide (H 2 S) donor. FW1256 inhibits NF-κB activity and induces cell apoptosis. FW1256 exerts potent anti-inflammatory effects and has the potential for cancer and cardiovascular disease treatment. In Vitro FW1256 (200 µM; 24.5 hours; AW264.7 cells) treatment significantly reduces IL-1β, COX-2 and iNOS mRNA and protein in LPS-stimulated RAW264.7 macrophages. FW1256 (200 µM; 24.5 hours; AW264.7 cells) treatment significantly reduces IL-1β, COX-2 and iNOS PROTE and protein in LPS-stimulated RAW264.7 macrophages. FW1256 concentration dependently decreases TNF-α (IC 50 of 61.2 µM), IL-6 (IC 50 of 11.7 µM), PGE2 (IC 50 of 25.5 µM) and NO (IC 50 of 34.6 µM) generation in LPS-stimulated RAW264.7 macrophages and bone marrow-derived macrophages (BMDMs) (IC 50 s of 414.9 µM, 300.2 µM, 4 µM and 9.5 µM for TNF-α, IL-6, PGE2 and NO, respectively). FW1256 decreases NF-κB activation as evidenced by reduced cytosolic phospho-IκBα levels and reduces nuclear p65 levels in LPS-stimulated RAW264.7 macrophages treated with FW1256. MCE has not independently confirmed the accuracy of these methods. They are for reference only. RT-PCRCell Line: RAW264.7 cells Concentration: 200 µM Incubation Time: 24.5 hours Result: Significantly reduced IL-1β, COX-2 and iNOS mRNA in LPS-stimulated RAW264.7 macrophages.. Western Blot AnalysisCell Line: RAW264.7 cells Concentration: 200 µM Incubation Time: 24.5 hours Result: Significantly reduced IL-1β, COX-2 and iNOS proteinin LPS-stimulated RAW264.7 macrophages.. In Vivo FW1256 (100 mg/kg; intraperitoneal injection; male C57BL/6 mice) treatment reduces IL-1β, TNFα, nitrate/nitrite and PGE2 levels in LPS-treated mice . MCE has not independently confirmed the accuracy of these methods. They are for reference only. Animal Model: Male C57BL/6 mice (20-25 g, 6-10 weeks) injected with E. coli lipopolysaccharide (LPS) Dosage: 100 mg/kg Administration: Intraperitoneal injection Result: Reduced IL-1β, TNFα, nitrate/nitrite and PGE2 levels in LPS-treated mice. Form:Solid