Figure 2
Figure 2. P17-induced adhesion and migration of monocytes involve a PTx-sensitive and Rho/Rock–dependent signaling pathway. (A-B) Static adhesion of monocytes on fibrinogen. Monocytes pretreated with 500 ng/mL of PTx for 2 hours at 37°C were stimulated for 2 minutes at 37°C with PBS (NT), GST, fMLP, p17 (A) or for 10 minutes with phorbol 12-myristate 13-acetate (PMA; B). Bars represent the means ± SD of 3 independent experiments performed in triplicate. Statistical analysis was performed by paired 2-tail Student t test. ***P < .001. (C) Transwell migration assay of monocytes in response to the indicated treatments. Monocytes pretreated for 2 hours at 37°C with 500 ng/mL of PTx were stimulated for 90 minutes at 37°C with PBS (NT), GST, L-α-lysophosphatidylcholine palmitoyl C16:0 (LPC), fMLP, or p17. Bars represent the means ± SD of 3 independent experiments performed in duplicate. Statistical analysis was performed by paired 2-tail Student t test. ***P < .001. (D) Static adhesion of monocytes on fibrinogen. Monocytes were pretreated for 20 minutes at 37°C with 25μM of the ROCK inhibitor Y27632 and then stimulated for 2 minutes at 37°C with PBS (NT), GST, fMLP, or p17. Bars represent the means ± SD of 3 independent experiments performed in triplicate. Statistical analysis was performed by paired 2-tail Student t test. *P < .05. (E) Transwell migration assay of monocytes in response to the indicated treatments. Monocytes pretreated for 20 minutes at 37°C with 25μM of the ROCK inhibitor Y27632 were stimulated for 90 minutes at 37°C with PBS (NT), GST, fMLP, or p17 (50nM). Bars represent the means ± SD of 3 independent experiments performed in duplicate. Statistical analysis was performed by paired 2-tail Student t test. ***P < .001.

P17-induced adhesion and migration of monocytes involve a PTx-sensitive and Rho/Rock–dependent signaling pathway. (A-B) Static adhesion of monocytes on fibrinogen. Monocytes pretreated with 500 ng/mL of PTx for 2 hours at 37°C were stimulated for 2 minutes at 37°C with PBS (NT), GST, fMLP, p17 (A) or for 10 minutes with phorbol 12-myristate 13-acetate (PMA; B). Bars represent the means ± SD of 3 independent experiments performed in triplicate. Statistical analysis was performed by paired 2-tail Student t test. ***P < .001. (C) Transwell migration assay of monocytes in response to the indicated treatments. Monocytes pretreated for 2 hours at 37°C with 500 ng/mL of PTx were stimulated for 90 minutes at 37°C with PBS (NT), GST, L-α-lysophosphatidylcholine palmitoyl C16:0 (LPC), fMLP, or p17. Bars represent the means ± SD of 3 independent experiments performed in duplicate. Statistical analysis was performed by paired 2-tail Student t test. ***P < .001. (D) Static adhesion of monocytes on fibrinogen. Monocytes were pretreated for 20 minutes at 37°C with 25μM of the ROCK inhibitor Y27632 and then stimulated for 2 minutes at 37°C with PBS (NT), GST, fMLP, or p17. Bars represent the means ± SD of 3 independent experiments performed in triplicate. Statistical analysis was performed by paired 2-tail Student t test. *P < .05. (E) Transwell migration assay of monocytes in response to the indicated treatments. Monocytes pretreated for 20 minutes at 37°C with 25μM of the ROCK inhibitor Y27632 were stimulated for 90 minutes at 37°C with PBS (NT), GST, fMLP, or p17 (50nM). Bars represent the means ± SD of 3 independent experiments performed in duplicate. Statistical analysis was performed by paired 2-tail Student t test. ***P < .001.

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