Dose-dependent induction and kinetics of CCR6 mRNA expression in cytokine-activated PMNLs.
(A) PMNLs were cultured with increasing concentrations of TNF-α (0-10 ng/mL), IFN-γ (0-25 U/mL), or both for 6 hours. (B) PMNLs were cultured with increasing concentrations of TNF-α (0-10 ng/mL), IFN-γ (0-0.25 U/mL), or both in the presence of GM-CSF (0.05 ng/mL). (C) PMNLs were cultured in the presence or absence of TNF-α (1 ng/mL), IFN-γ (2.5 U/mL), or both for 1, 3, 6, and 16 hours. Total RNA was extracted and subjected to Northern blot analysis. The blots were hybridized with 32P-labeled human CCR6 or β-actin cDNA probe. Autoradiographic signals were quantified, standardized against the levels of β-actin, and presented as relative density. The level of expression induced by 1 ng/mL TNF-α equals 1 in the chart. Representative of several experiments with similar results.
