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. 2010 Jul;24(4):308–316. doi: 10.1016/S1674-8301(10)60043-7

Fig.2. Effect of IL-13 on LPS-induced NF-κB activation.

Fig.2

A. A competitional gel shift assay for NF-κB DNA-binding activity. EMSA with nuclear extracts from unstimulated or LPS-activated HMCs was carried out. 100-fold molar excess unrelated oligonucleotide containing the Sp-1 binding motif (cold Sp-1) did not inhibit binding of the radiolabeled NF-κB oligonucleotide, whereas 100-fold molar excess unlabeled NF-κB consensus sequence (cold κB) completely abrogated the signal. B: Supershift assay showed the interaction of anti-p65, anti-p50 and anti-cRel antibodies with NF-κB protein stimulated by LPS. C: HMCs were pretreated at 37°C for 30 min with different concentrations (0-100 ng/mL) of IL-13 followed by 1 h incubation with 10 µg/mL LPS. After these treatments, nuclear extracts were prepared and then assayed for NF-κB as described in materials and methods. D: Quantitation of EMSA blots by image analysis (C) was expressed in arbitrary units as the ratio of the control. Values represent mean±SD (n = 3, **P < 0.01).