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Figure 4 | BMC Molecular Biology

Figure 4

From: Interleukin-4 inhibition of interleukin-1-induced expression of matrix metalloproteinase-3 (MMP-3) is independent of lipoxygenase and PPARγ activation in human gingival fibroblasts

Figure 4

A specific inhibitor of 5-lipoxygenase, MK886, super-induces MMP-3 mRNA and protein. HGF cultures were pre-treated with NDGA at the indicated doses for 1 hour prior to addition of IL-1 (10 ng/ml) and/or IL-4 (10 ng/ml). A) Total RNA was harvested 12 hours after cytokine treatment, quantitated by real-time PCR and normalized to levels of GAPDH mRNA. The graph represents five independent experiments performed in triplicate, expressed as mean +/- SEM. B) Conditioned medium was collected from cultures after 24 hours incubation with cytokines +/- MK886 at the indicated doses. MMP-3 protein levels were determined by ELISA. The graph shows data from two different HGF cultures derived from two different donors. C) COS-1 cells were transiently transfected with a luciferase reporter construct containing 2300 bp of the human MMP-3 5' flanking sequence, along with an SV-β-gal reporter as a control for transfection efficiency. MK886 was added 16 hours after transfection, and cells were harvested 24 hours later. Shown are data from three independent experiments performed in triplicate, normalized to levels of β-galactosidase, and expressed relative to control, mean +/- SEM. Statistical significance was determined using Student's t-test.

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