Fig. 8
From: Network inference from glycoproteomics data reveals new reactions in the IgG glycosylation pathway
Experimental validation results. a In vitro enzymatic assay. The figure illustrates the ratio of G1FS1 over G2FS1 across different concentrations of the enzyme (B4GalT1), and in presence or absence of sugar donors. Bars represent the average value over triplicates, while error bars represent standard deviations. With increasing enzyme concentrations, the glycan ratio decreases significantly with respect to the corresponding negative control, confirming the occurrence of the predicted reaction. P-values were obtained from a two sample t-test. b Quantitative overlap between the localization of the three enzymes. The overall colocalization of each enzyme pair is expressed as an overlap coefficient percentage (mean % ± standard deviation). We observe substantial colocalization of all enzyme pairs in both cell lines. c Exemplary colocalization images of B4GalT1, ST6Gal-1, and MGAT3 in CaCo-2 (left) and COS-7 (right) cells, used for the overlap quantification in b. The individual figures represent a typical view from five different Golgi areas examined. In the images labeled as “Merged” and “Zoom”, yellow areas represent enzymatic overlap. Due to the dispersed Golgi stacks throughout the cytoplasm in Caco-2 cells, the overlap can be observed clearly in separated cisternae, proving that localization of the glycosyltransferases is not limited to cis-Golgi, medial-Golgi, or trans-Golgi areas. Bar represents 5 μm
