Evidence-Based Complementary and Alternative Medicine / 2019 / Article / Fig 2

Research Article

Anti-Inflammatory Effects of Licania macrocarpa Cuatrec Methanol Extract Target Src- and TAK1-Mediated Pathways

Figure 2

Anti-inflammatory effects of Lm-ME on mRNA expression and transcriptional activity. (a) RAW264.7 cells were pretreated with 50, 100, or 200 μg/ml of Lm-ME and then treated with 1 μg/ml of LPS. After 6 h, mRNA was extracted from the cells, and proinflammatory cytokines (iNOS, COX-2, IL-6, and IL-1β) were evaluated using semiquantitative RT-PCR. (b–e) In 24 wells, HEK293T cells were plated at 1.25 × 105 cells per well. After 24 h, NF-κB or AP-1 luciferase gene and β-galactosidase-expressing constructs were transfected into HEK293T cells along with either the MyD88 or TRIF gene. After 24 h incubation, 50, 100, or 200 μg/ml of Lm-ME was added, and NF-κB luciferase activity was measured by the luminometer. Total form of FLAG and TRIF was determined by western blotting analysis. (d) RAW 264.7 cells were pretreated with 200 μg/ml of Lm-ME for 30 min and then exposed to 1 μg/ml LPS for 15, 30, or 60 min. Using western blotting analysis, nuclear c-Jun, and c-Fos levels were determined.

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