IL-1β induces translational up-regulation of egr2. (A) Secretion of IL-1β in Ctr and CM was analyzed using CBA assays (n > 3). (**) P < 0.01). (B) CM was pretreated with IgG control (5 μg/mL) or neutralizing IL-1β antibody (5 μg/mL) for 1 h at 37°C. Then, MCF7 cells were incubated with the respective CMs for 4 h, followed by polysomal fractionation as described before. Changes of egr2 mRNA in pooled polysomal fractions 6–10 in response to CM+αIL-1β relative to CM+IgG were determined. (C) MCF7 cells were treated with human recombinant IL-1β (50 ng/mL) for 4 h, followed by polysomal fractionation as described before. IL-1β-induced changes for egr2 mRNA in pooled polysomal distribution relative to control were analyzed using RT-qPCR. Data are normalized to gapdh and presented as means ± SEM (n ≥ 3). (*) P < 0.05.
