Up-regulation of CCR6 mRNA expression in PHA-sup–stimulated or cytokine-stimulated PMNLs.
(A) PMNLs were cultured with medium alone, PHA (2.5 μg/mL), or PHA-sup in the presence or absence of neutralizing IgG (5 μg/well) against TNF-α, IFN-γ, or GM-CSF, for 6 hours. (B) PMNLs were cultured with LPS (1 ng/mL), TNF-α (1 ng/mL), IFN-γ (2.5 U/mL), GM-CSF (0.05 ng/mL), or IL-1β (0.05 ng/mL), rTNF-α (1 ng/mL), and IFN-γ (2.5 U/mL), TNF-α (1 ng/mL), and GM-CSF (0.05 ng/mL), or IFN-γ (2.5 U/mL) and GM-CSF (0.05 ng/mL) for 6 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 and exposed to x-ray film. Data were also analyzed by densitometry. Representative of several experiments with similar results.
