Figure 10.
GNE activity resulted in decreased galectin-1 binding. Cells were cultured for 24 h in high glucose media (A), low or high glucose media (B), high glucose media supplemented with 100 μm Ac4GlcNAc (C), or high glucose media supplemented with free 40 mm GlcNAc (D). Binding of 3 μm biotinylated galectin-1 was measured using an ELISA assay and normalized to total protein content. Cell lines labeled in red did not express an active GNE epimerase domain and did not synthesize sialic acid. Data shown represent three or four biological replicates with error bars depicting the mean and S.E. Each experiment was performed at least twice. Statistical significance determined by unpaired Welch's test: *** indicates a p value < 0.001, ** indicates a p value < 0.01, * indicates a p value < 0.05. ns indicates difference not statistically significant.