Myelin Induces Inflammation Figure 8. Effect of myelin on activation of PI3K/Akt pathway. (A) Phosphorylation of p85 and Akt in bone marrow-derived macrophages from WT and CR3 KO mice treated with myelin for the indicated time. (B) Effect of PI3K inhibitor LY-294002 on myelin-induced IkB-a phosphorylation and degradation in macrophages. (C) Effect of Akt inhibitor IV on myelin-induced IkB-a phosphorylation and degradation in macrophages. (D) Effect of inhibitors of PI3K, Akt, NF-kB and PKC on myelin-induced Akt phosphorylation in macrophages. (E) Macrophages were pretreated with inhibitors of PI3K, Akt, NF-kB and PKC for 30 minutes, respectively and then incubated with myelin for 90 minutes. Myelin-induced p65 translocation in macrophages was detected by immunofluorescent staining. p65 was stained red and nuclei were stained with DAPI (blue) (Scale bar = 10 mm). All inhibitors were used for 30-minute pre-incubation before myelin incubation. Phosphorylation levels were normalized to non-phosphorylated total proteins. Similar results were obtained from three independent experiments. doi:10.1371/journal.pone.0009380.g008 PLoS ONE | www.plosone.org 11 February 2010 | Volume 5 | Issue 2 | e9380