Fig. 3.

Azithromycin (AZM) inhibits nuclear translocation of nuclear factor kappa B (NF-κB). (a) Electrophoretic mobility shift assay (EMSA) showing the binding of nuclear factors to the NF-κB p65 wild-type oligo probe. The labelled NF-κB probe was incubated with nuclear extract from immature DCs (im-DCs) stimulated with or without lipopolysaccharide (LPS). In lane 1, a labelled NF-κB probe was added to control im-DCs. In lanes 2 or 3, a labelled NF-κB probe with a 100-fold molar excess of unlabelled NF-κB wild-type or mutant competitor oligos was added to im-DCs stimulated with LPS, respectively. In lane 4, labelled NF-κB probe was added to control im-DCs stimulated with LPS. In lanes 5–7, a labelled NF-κB probe was added to AZM-treated im-DCs stimulated with LPS. In lane 5, 50 µg/ml AZM was added on day 6. In lane 6, 75 µg/ml AZM was added on day 6. In lane 7, 50 µg/ml AZM was added on days 0, 3 and 6. (b) The NF-κB DNA–protein complexes were analysed on a Fuji BASS 1000 imaging analyser. The results are representative of three independent experiments. n.s.: not significant' *P < 0·01; **P < 0·001.