Fig. 1.

SOD3 is released from a preformed pool. HMDMs were stimulated by the addition of LPS and the supernatant collected at the indicated time points. The supernatant and corresponding cell lysates were analyzed by (A) immunoprecipitation/Western blotting. As a control of immunoprecipitation, we performed parallel analysis in the absence (−) or presence (+) of purified recombinant SOD3. Additionally, the presence of SOD3 in the supernatant was evaluated by ELISA (B). (C) The transcriptional level of SOD3 upon LPS stimulation was evaluated by RT-qPCR and presented relatively to the level in the absence of LPS (t = 0 min). This analysis shows that the release of SOD3 from macrophages is relative fast and does not reflect a transcriptional upregulation. Error bars in panel B and C represents mean ± SD (n = 3).