Expression of GlcNAc6ST-2 in cultured HUVEC. (a) Real time RT-PCR quantification of GlcNAc6ST-2 mRNA in cultured HUVEC. HUVEC cultures were stimulated for 8 h with LT-αβ (LT), TNF-α (TNF), or interferon-γ (IFN) and mRNA obtained from stimulated and non-stimulated cells. Basal (non-stimulated) expression was set to 1 and the GlcNAc6ST-2/β-actin ratio in stimulated cells is shown (mean ± SD of three independent experiments, each including triplicate HUVEC cultures). (*) p < 0.05. (b) Western blot analysis of GlcNAc6ST-2 in cultured HUVEC stimulated for 24 h with LT-αβ (LT) or TNF-α (TNF), (data are representative of three independent experiments with different HUVEC lines).(c) Immunofluorescent detection of GlcNAc6ST-2 protein in cultured HUVEC endothelial cells non-stimulated (Basal) or stimulated for 24 h with LT-αβ (LT) or TNF-α (TNF). In control (CTRL) panel, anti-GlcNAc6ST-2 antibody was replaced by non-immune rabbit IgG (images are representative of three independent experiments).