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Figure 4 | Journal of Inflammation

Figure 4

From: 15-deoxy-delta12,14-prostaglandin J2 attenuates endothelial-monocyte interaction: implication for inflammatory diseases

Figure 4

15d-PGJ2 inhibits TNFα induced NF-κB function in endothelial cells. bEND.3 cells were treated with 15d-PGJ2 (1–10 μM) and TNFα (50 ng/ml) for various time periods (5–40 min) and processed for EMSA as described in 'Material and Methods' (A). bEND.3 cells were transiently transfected with p65, p50 expression vectors along with NF-κB luciferase reporter construct (0.5 μg/well) and pCMV-β-galactosidase (0.5 μg/well) followed by treatment with 15d-PGJ2 (5–20 μM) for 4 h and processed for luciferase and β-galactosidase activities. Luciferase activity was normalized with respect to β-gal activity (B). Results were calculated as mean ± SD for 3 independent experiments. Samples were run in triplicates. &&& p < 0.001 compared with control and !!! p < 0.001 compared with TNFα treatment (50 ng/ml). Cells were treated with 15d-PGJ2(10 μM) for 30 min followed by TNFα for 20 min and stained with anti-p65 antibody as described in 'Material and Methods' (C). Images taken at 200× magnification are representative of 6 fields from each treatment and 3 independent experiments. Treated and untreated cells were processed for immunoblot analysis for p65 and IκBα levels (D). Representative blot from two independent experiments are shown.

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