CM induces egr2 translation in a p38-MAPK-dependent manner. (A) MCF7 cells were treated with human recombinant IL-1β (50 ng/mL) to the indicated time points. Whole-cell extracts were subjected to Western analysis and probed with the indicated antibodies. Blots are representative for at least three independent experiments. (B) MCF7 cells were treated with CM or Ctr to the indicated time points. Whole-cell extracts were subjected to Western analysis and probed with the indicated antibodies. Blots are representative for at least three independent experiments. (C) MCF7 cells were treated for 4 h with CM alone or in combination with SB203580 (10 μM), followed by polysomal fractionation. Change of egr2 mRNA in pooled polysomal fractions 6–10 in response to CM+SB is given relative to CM. Data are normalized to gapdh and presented as means ± SEM (n ≥ 3). (**) P < 0.01. (D) MCF7 cells were treated for 15 min with Ctr and CM pretreated with IgG (5μg/mL) or IL-1β neutralizing antibody (5μg/mL) as described before. Whole-cell extracts were subjected to Western analysis and probed with the indicated antibodies, followed by densitometric analysis. Levels of phospho-p38 were normalized to total p38 and are presented relative to Ctr. Blots are representative for at least three independent experiments.
