FIGURE 7.

Src phosphorylation leads to its subcellular redistribution. A, MMCs were treated with poly(I:C) for 1 h and the levels of pY⁴¹⁶Src, Src, and actin in the 0.5% Triton X100-soluble fraction were analyzed by Western Blot. B, MMC lysates were prepared by solubilizing in 1% SDS containing buffer followed by sonication, the levels of Src and actin in cell lysates were analyzed by Western Blot. C, Wt11 cells were treated with poly(I:C) for 4 h, the raft fractions and the soluble fractions were separated by sucrose-gradient ultracentrifugation and analyzed for Src levels by Western Blot. Flotillin-1 was used as a marker for lipid raft fractions. D, Wt11 cells were incubated with poly(I:C) for the indicated time (right), the cell extracts were analyzed by sucrose gradient ultracentrifugation that separates lipid rafts from soluble fractions; the gradient fractions (1.2 ml each) were collected from the top to the bottom of ultracentrifuge tube, and subjected to Western Blot for Src, Flotillin-1 and FAK (a soluble fraction marker). E, MMCs were treated with poly(I:C) for 4 h, cell lysates were immunoprecipitated with Cbp/PAG and the immunoprecipitates were analyzed for Src by Western Blot.