American Society of Hematology

Fig. 4.

$Fig. 4. (A) Inhibition by sphingosine of the translocation of Raf-1 to the plasma membrane during phagocytosis of EIgG in PMN. PMN (1 × 108/mL) were incubated with EIgG (5 × 109/mL) at 37°C. The PMN were then separated into cytosolic and membrane fractions. The fractions were analyzed for the presence of Raf-1 by employing SDS-PAGE and immunoblotting using specific Abs. (A) indicates the translocation of Raf-1 to the plasma membrane within 3 minutes in column B, a decrease in translocation at 20 minutes phagocytosis in column C, respectively; columns D indicate PMN pretreated 10 μmol/L sphingosine and activated with EIgG for 3 minutes. (B) Restoration of PKCδ translocation from cytosol to the plasma membrane in PMN treated with sphingosine by DiC10. (B) indicates translocation of PKCδ to the plasma membrane at 3 minutes from the cytosol in the presence or in the absence of 10 μmol/L sphingosine and DiC10.$

(A) Inhibition by sphingosine of the translocation of Raf-1 to the plasma membrane during phagocytosis of EIgG in PMN. PMN (1 × 10⁸/mL) were incubated with EIgG (5 × 10⁹/mL) at 37°C. The PMN were then separated into cytosolic and membrane fractions. The fractions were analyzed for the presence of Raf-1 by employing SDS-PAGE and immunoblotting using specific Abs. (A) indicates the translocation of Raf-1 to the plasma membrane within 3 minutes in column B, a decrease in translocation at 20 minutes phagocytosis in column C, respectively; columns D indicate PMN pretreated 10 μmol/L sphingosine and activated with EIgG for 3 minutes. (B) Restoration of PKCδ translocation from cytosol to the plasma membrane in PMN treated with sphingosine by DiC10. (B) indicates translocation of PKCδ to the plasma membrane at 3 minutes from the cytosol in the presence or in the absence of 10 μmol/L sphingosine and DiC10.

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