Expression of ENA-78 and CXCR-2 and chemotaxis of neutrophils.

(A) mRNA expression of ENA-78 in neutrophils treated with or without G-CSF. Total RNAs were obtained from the untreated neutrophils and those treated with 10 ng/mL G-CSF for 24 hours and then used for Northern blot analysis. Representative result of 3 different experiments is shown. (B) Expression of CXCR-2 on neutrophils. The untreated neutrophils and those treated with 10 ng/mL G-CSF for 24 hours were stained with anti–CXCR-2 antibody and fluorescein isothiocyanate–conjugated second antibody and then analyzed for expression of CXCR-2 with a FACScaliber. Representative result of 2 different experiments is shown. (C) Chemotaxis of neutrophils stimulated by conditioned medium of neutrophils. Supernatants of neutrophils treated with or without G-CSF (G⁺ or G⁻, respectively) at 10 ng/mL were placed at 20% in the lower chambers. Anti–ENA-78 or anti–CXCR-2 antibody was added in the lower chambers at 2 μg/mL. Recombinant ENA-78 (10 ng/mL) was used as a positive control. Results are presented as percentage of numbers of transmigrated neutrophils treated with G CM. Data present the mean ± SD of 3 different experiments.