Fig. 7.

HLF-derived PGs and PG metabolites activate PPARγ and inhibit NFκB. A: PGs and PG metabolites identified in the lipidome of activated HLFs [or vehicle (0.1% DMSO)] were applied at 5 μM final concentration to a PPRE-luciferase reporter in the presence of the PPARγ inhibitor GW-9662 (10 μM) or vehicle (0.1% DMSO); rosiglitazone (50 nM) and 15d-PGJ₂ (1 μM) were used as positive controls. After 16 h of incubation, the ratio of firefly to Renilla luciferase activity was calculated in the presence and absence of GW-9662 and normalized as described in materials and methods. B: PGs and PG metabolites identified in the lipidome of activated HLFs [or vehicle (0.1% DMSO)] were applied at 5 μM final concentration to a NFκB-response element-luciferase reporter for 1 h prior to addition of IL-1β (10 pg/ml) or vehicle (PBS), and reporter cells were incubated for an additional 16 h prior to determination of firefly and Renilla luciferase levels as described in materials and methods. The NFκB inhibitors SC-514 (20 μM) and BAY 11-7082 (1 μM) were used as controls. Values are means ± SD of 3 independent biological replicates per condition. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001 vs. vehicle alone (by unpaired t-test).