Figure 1

LPS stimulates Shc protein accumulation and activation in bone marrow-derived DCs. Purified DCs were stimulated with LPS (0.5 μg/ml), and Shc protein accumulation was measured at 48 h (A). For Shc activation, DCs were treated with LPS (0.5 μg/ml) with mouse control IgG or anti-Shc mAb (2 μg/ml) for 10 or 30 min, or with LPS (0.5 μg/ml) in the presence of 10 nM SU6656 and/or mouse control IgG (2 μg/ml) for 10 min, as indicated (B). Cell lysate (30 μg) from each sample was subjected to Western blot with anti-Shc, anti-phosphotyrosine 317-Shc (pTyr317-Shc) and anti-phosphotyrosine 239/240-Shc (pTyr239/240-Shc) mAb. The bound antibodies were detected by the ECL system. The significant change of pTyr239/240-Shc was observed and the statistics at 10-min was presented. The results are representative of three independent experiments. (n = 3; **, P < 0.01)