Figure 6

Activation of Src and PKCδ by daidzein treatment and suppression of the PKCδ activation by the specific Src inhibitor PP2. (A) Phosphorylation of Src following daidzein treatment was analyzed by Western blot analysis. DRG neuronal cultures were treated with 30 μM daidzein for 0, 15, 30, or 60 min, then the cell homogenate was analyzed for phosphorylated Src (pSrc416) and total Src. Upper panel showed a representative blot from one experiment. Lower panel showed optical densities of the densitometric scans of the pSrc416 bands. Basal levels of phosphorylation in non-stimulated cells (DMSO) were taken as 100% for each individual treatment. *, p < 0.05 vs 0 min, n = 4. (B) Phosphorylation of PKCδ was analyzed by Western blot analysis. DRG neuronal cultures were given DMSO, 30 μM daidzein (Dz) for 30 min, 10 μM the Src inhibitor PP2 for 30 min followed by 30 μM daidzein for 30 min (Dz + PP2), or 10 μM PP2 for 60 min (PP2). The cell homogenate was analyzed for phosphorylated and total PKCδ. Upper panel showed a representative blot from one experiment. Lower panel showed optical densities of the densitometric scans of the PKCδ bands. PP2 treatment reduced both basal and. daidzein-induced PKCδ phosphorylation. *, p < 0.05 vs DMSO; #, p < 0.05 vs daidzein. n = 4.