Effect of IL-15 and IL-2 towards the activation of nuclear AP-1 DNA-binding activities in human peripheral blood neutrophils and lymphocytes. (A) Neutrophils or autologous PBL were cultured for 15 minutes in the presence or absence of 250 ng/mL IL-15 (ie, 20 nmol/L) or 103 U/mL IL-2 (∼5 nmol/L); nuclear extracts were then prepared and analyzed in EMSA using a consensus AP-1 oligonucleotide probe. The amount of extract used in the binding reactions corresponded to 5 μg protein for neutrophils, and to 2 μg protein for PBL. This experiment is representative of three (neutrophils) and four (PBL). (B) Nuclear extracts from IL-15–activated PBL (1.5 μg of protein) were prepared as described in (A), and incubated without antibodies (–), or with specific antibodies to c-jun, junB, junD, c-fos, and fosB, before the addition of labeled AP-1 probe and subsequent EMSA analysis. This experiment is representative of three. Closed arrowheads indicate the major inducible AP-1 complex; open arrowheads indicated supershifted bands.
