Induction of arginase-1 expression by PLT-sup in BMDMs.

<p>(A) The gene expressions of <i>Tnf</i>, <i>Il6</i>, <i>Il1b</i>, <i>Nos2</i>, <i>Arg1</i>, <i>Fizz1</i>, <i>Ym1</i>, and <i>Mrc1</i> in PLT-BMDMs and control BMDMs were analyzed by RT-qPCR with the relative standard curve method using <i>Gapdh</i> as an internal control. The gene expression in PLT-BMDMs was represented as the value relative to gene expression in control BMDMs. Experiments were performed in triplicate and repeated three times. Data are presented as the mean ± SEM, and representative results of the three experiments are shown. (B) PLT-BMDMs and control BMDMs were lysed in 1 × SDS sample buffer, and the resultant lysates were subjected to western blotting analysis with antibodies against arginase-1 or GAPDH. The relative intensity of each arginase-1 band after normalization to the corresponding levels of GAPDH is shown above each blot. Experiments were repeated five times, and representative results are shown. (C) PLT-BMDMs and control BMDMs (4 × 10⁵ cells) were lysed with 0.1% Triton X-100 (100 μL) containing a protease inhibitor cocktail for 10 min at room temperature, and the lysate was assayed for arginase activity as described in the Materials and Methods. Experiments were performed in quintuplicate and repeated five times. Data are presented as the mean ± SEM. **p < 0.01 vs. control BMDMs. Representative results of the five experiments are shown.