Figure 2.
The effect of IL-10 on NF-kB activation is mediated through PI3K and AKT. (A) Addition of recombinant IL-10 protein to cultured spinal cord neurons in vitro induced phosphorylation of Stat3, AKT (S473), IKK (S180/181) and GSK3β (S9). Western blot of cultured spinal cord neuron lysates after 5, 10, 20, 40 min and 12 h of IL-10 treatment (10 ng/ml). (B) The effects of IL-10 on phosphorylation of AKT, IKK and GSK3β and on IκBα levels were blocked by LY294002. (C) Increases in p50 and p65 NF-κB in the nuclear fraction of cultured spinal cord neurons induced by IL-10 was partially blocked by LY294002. Histone 3 is used as an internal control for the nuclear fraction. Cultured cells were treated with 50 μM LY294002 for 30 min, followed by exposure to IL-10 (10 ng/ml) for 40 min (B) and 12 h (C). Quantitative analysis of the blot results is presented in Supplementary Figures 2 and 3.