Figure 10.

Effect of Oga knockdown on LPS-mediated iNOS/NO induction. Oga knockdown cells were generated via stable transfection of shCont, shOga1, and shOga2. Control and Oga knockdown cells were stimulated with LPS (100 ng/ml) with or without 5 mm GlcN for 24 h. Whole cell lysates were prepared and the expression levels of OGA, O-GlcNAc, iNOS, and OGT were measured via Western blotting using anti-MGEA5, CTD110.6, anti-iNOS, and anti-OGT antibodies, respectively (A). Relative densitometric intensities were quantitatively measured and normalized to GAPDH levels (B). Nitrite levels in cell culture medium were measured with the Griess assay (C). Blots are representative of three independent experiments. All values are presented as mean ± S.E. Asterisks denote significantly increased from the untreated control (*, p < 0.05; **, p < 0.01); hash mark (#) indicates significantly decreased from LPS-stimulated conditions (p < 0.05); &, indicates significantly increased from shCont LPS-stimulated conditions (p < 0.05).