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Fig. 2 | Cell Communication and Signaling

Fig. 2

From: ARF1 recruits RAC1 to leading edge in neutrophil chemotaxis

Fig. 2

Requirement of GBF1 in RAC1 activity and its translocation to the leading edges. (a-f) Subcellular localization of CDC42, RAC1, or RAC2. Differentiated HL-60 cells, transfected with GBF1 siRNA or an irrelevant RNA duplex (Irr), were incubated with or without fMLP for 15 min, and subjected to anti-CDC42 (a), anti-RAC1 (c), or anti-RAC2 immunostaining (e), and percentages of CDC42 molecules (b), RAC1 molecules (d), and RAC2 molecules (f) translocated to the leading edge in fMLP-stimulated cells were calculated. F-actin was visualized by Texas Red-phalloidin. Data are representative images of three independent experiments (a, c, and e), and >25 cells were analyzed in three independent experiments (b, d, and f). (g and h) Activities of CDC42, RAC1, and RAC2. Activities of CDC42, RAC1, and RAC2 were measured by GST-PBD pulldown coupled with the indicated antibodies. Each lower panel represents immunoblots of total cell lysates (5 μg) by the indicated antibodies. Data are representative of three independent experiments (g), and were analyzed in three independent experiments (h). In h, values for Irr control at 0 s are considered 1. Error bars, SEM (b, d, f and h). ** represents a statistical difference from the Irr control (p < 0.01). Bars, 10 μm (a, c, and e)

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