The mouse macrophage cell line RAW264.7 was purchased from ATCC and incubated in RPMI 1,640 with 10% fetal bovine plasma in a 37°C sterile incubator with 5% CO2. Cells were seeded in 6- or 24 -well plates and exposed to TPPU (1 μM, Cayman Chemical, Ann Arbor, Mich) or 14,15-EET (1 μM, Cayman Chemical, Ann Arbor, Mich) for 4 h after they reached 80% to 90% confluence. LPS (1 μg/mL) was then added and co-incubated for 12 h. 14,15-EEZE (an inhibitor of 14,15-EET, 1 μM, Cayman Chemical, Ann Arbor, Mich) was added 1 h before LPS stimulation. Cell supernatants were collected for inflammatory cytokines detection (20 (link)).
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Macrophage Modulation by EET and TPPU
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Corresponding Organization : Tongji Hospital
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Variable analysis
independent variables
- TPPU (1 μM)
- 14,15-EET (1 μM)
- 14,15-EEZE (an inhibitor of 14,15-EET, 1 μM)
- Inflammatory cytokines detection
- Cells were incubated in RPMI 1,640 with 10% fetal bovine plasma in a 37°C sterile incubator with 5% CO2
- Cells were seeded in 6- or 24 -well plates
- Cells were exposed to the independent variables for 4 h after they reached 80% to 90% confluence
- LPS (1 μg/mL) was then added and co-incubated for 12 h
- 14,15-EEZE was added 1 h before LPS stimulation
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