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. 2009 Jul 20;86(4):833–845. doi: 10.1189/jlb.0908551

Figure 3.

Figure 3.

MIP-1β-induced chemotaxis of macrophages requires class IA PI3K activation. (A) Macrophages were pretreated for 1–2 h with the PI3K inhibitor LY294002 (10 μM), wortmannin (100 nM), or control vehicle alone (0.1% DMSO) prior to exposure to MIP-1β. (B) Chemotaxis of unstimulated (open bars) or MIP-1β-stimulated (solid bars) macrophages that had been transfected with control or PI3Kp85-specific siRNA. Inset, Immunoblots for Pyk2, PI3Kp85, PI3Kp101, Lyn, and β-actin protein were performed in parallel on the same batch of siRNA-transfected macrophages. (C) Macrophages were pretreated for 30–60 min with the isoform-specific class IA PI3K inhibitors (horizontal striped bars) PI-103 (40 nM) and PI3Kαi2 (10 nM), class IB PI3K inhibitors (vertical striped bars) AS605240 (10 nM) and AS604850 (250 nM), or control vehicle alone (open bar; 0.1% DMSO), followed by stimulation with MIP-1β. Data shown are means ± se of three independent experiments with cells from different donors, along with representative immunoblots (***, P<0.001).