MALP-2 stimulates translocation of Nrf2 to the nucleus and binding to HO-1 ARE in THP-1 cells.

<p><b>A.</b> Immunofluorescence images of THP-1 cells pretreated with or without LY294002 (25 µM) and MyD88 siRNA in the presence of MALP-2 (5 ng/ml) for 2 h. Cells were fixed and then stained with 4′-6-diamidino-2-phenylindole or anti-Nrf2 antibody, followed by incubation with cy3-conjugated anti-rabbit antibody. Images were obtained using a confocal microscope. <b>B.</b> EMSA was performed using MALP-2-stimulated nuclear THP-1 extracts and biotinylated HO-1-ARE probe. The specificity of Nrf2 was verified by competition analysis with an excess of nonlabeled specific (cold probe) or nonspecific (NF-κB probe) oligonucleotide probe. For supershift analysis, nuclear extracts from MALP-2-treated THP-1 cells were preincubated with 1 µg anti-Nrf2 supershift antibody before EMSA.</p>