American Society of Hematology

Figure 4.

$Figure 4. PF4 induces protein translocation of small GTPases to the plasma membrane. (A-B) Neutrophils were stimulated with PF4 (4 μM) for the time periods indicated in the figure. Cells were lysed, and lysates were separated to obtain cytosol (cyt), Triton-insoluble membrane fraction (TI), raft-containing fractions (fractions 2-4), and non-raft membrane fractions (fractions 6-8). Fractions were separated on 10% SDS-PAGE, and Rac2 (A) and RhoA (B) were detected by Western blot analysis using specific antibodies. Bands were visualized by ECL. Data are representative of 3 independent experiments. (C-D) Neutrophils were stimulated with PF4 (4 μM) for 15 minutes or were left untreated, and GTPases were detected by indirect immunofluorescence staining with anti-Rac2 (C) or anti-RhoA (D) antibodies and were analyzed by confocal microscopy. Data from 1 of 3 representative experiments are given.$

PF4 induces protein translocation of small GTPases to the plasma membrane. (A-B) Neutrophils were stimulated with PF4 (4 μM) for the time periods indicated in the figure. Cells were lysed, and lysates were separated to obtain cytosol (cyt), Triton-insoluble membrane fraction (TI), raft-containing fractions (fractions 2-4), and non-raft membrane fractions (fractions 6-8). Fractions were separated on 10% SDS-PAGE, and Rac2 (A) and RhoA (B) were detected by Western blot analysis using specific antibodies. Bands were visualized by ECL. Data are representative of 3 independent experiments. (C-D) Neutrophils were stimulated with PF4 (4 μM) for 15 minutes or were left untreated, and GTPases were detected by indirect immunofluorescence staining with anti-Rac2 (C) or anti-RhoA (D) antibodies and were analyzed by confocal microscopy. Data from 1 of 3 representative experiments are given.

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