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Figure 5


Fig. 5. Phosphorylation of VASP upon CXCL8 stimulation is mediated through PKA and PKC. Lysates from HL-60-CXCR2 cells pretreated with various inhibitors, stimulated with vehicle (0 minutes) or 100 ng /ml CXCL8 for 1 or 5 minutes and analyzed by SDS-PAGE and western blot. Statistical difference of mean ± s.e.m. in DMSO versus treatment is indicated (*P≤0.05, Mann Whitney U-test). (A) Cells pretreated with DMSO or 20 µM H89 for 60 minutes and analyzed by western blot using antibodies specific for VASP and VASP Ser157-P. (B) Quantification of normalized density of VASP Ser157-P in DMSO-versus H89-treated samples. (C) Cells pretreated with DMSO or 5 µM rottlerin for 15 minutes and analyzed by western blot analysis for VASP and VASP Ser157-P. (D) Quantification of normalized density of VASP Ser157-P in DMSO-versus rottlerin-treated samples. (E) Cells pretreated with DMSO or 5 µM rottlerin for 15 minutes and analyzed for VASP and VASP Ser239-P (F). Quantification of normalized density of VASP Ser239-P in DMSO-versus rottlerin-treated samples. (G) Cells pretreated with DMSO or 20 µM H89 for 60 minutes and analyzed using antibodies for VASP and VASP Ser239-P. (H) Quantification of normalized density of VASP Ser239-P in DMSO-versus H89-treated samples. Data shown represent average quantification from at least three separate experiments.





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