Fig. 3.

AnxA1/ALX proresolving signaling signature leads to IL-10 release. (A) U937 cells (1 × 10⁶) or human primary monocytes (5 × 10⁵) were treated for 10 min with vehicle (Ctrl), AnxA1 (10⁻⁸ M) or SAA (10⁻⁷ M) and lysates subjected to Western blotting against phosphorylated and unphosphorylated p38, MAPKAPK1, MAPKAPK2, and Hsp27 (representative of three experiments). The cumulative data for Hsp27 are shown in bar graph (n = 3 distinct preparations). (B) Schema of the p38/MAPKAPK/Hsp27/IL-10 pathway. Human monocytes (1 × 10⁶) were treated with AnxA1 (10⁻⁸ M) for 6 h, with or without pretreatment with the p38 inhibitor SB203580 (10⁻⁷ M, 10 min). Data are mean ± SEM, three donors assayed in duplicate. **P < 0.01 vs. Ctrl, ⁺⁺⁺P < 0.001 vs. AnxA1 alone. (C) AnxA1 (1 μg) or saline (200 μL) were injected i.p. into wild-type or Alx-Fpr2/3 knockout (KO) mice and peritoneal lavages were harvested 6 h later. Data are mean ± SEM of six mice per group. *P < 0.05 vs. respective wild type. (D) Mice were treated with 10 mg/kg i.p. LPS and IL-10 levels were measured at the 24-h time point. (Inset) Endogenous AnxA1 levels in lavage fluids. Data are mean ± SEM of six mice per group. *P < 0.05 vs. wild type.