Figure 4.

Glycan biosynthesis and growth remain intact in B. fragilis and mammalian cells following treatment with S-glycosides. (A) Western blot analysis shows intact biosynthesis of polysaccharide A in B. fragilis treated with 0.1–2 mM thioglycosides BacSBn 4, DATSBn 5, and FucSBn 6 that is comparable to levels in samples treated only with Ac₄GalNAz. The increasing size of the shaded triangle corresponds to increasing concentrations of S-glycosides, with the short side of the triangle corresponding to 0.1 mM and the tallest part representing 2 mM. (B) B. fragilis exhibited robust growth across all treatments. (C) Western blot analysis of Ac₄GalNAz-labeled O-linked glycoproteins in AGS samples indicates that treatment with 10 μM thioglycosides BacSBn 4, DATSBn 5, and FucSBn 6 has no apparent effect on glycoprotein biosynthesis. Density (D) and viability (E) of AGS cells are not significantly impacted by thioglycoside treatment relative to untreated controls (−). These data are representative of replicate experiments (n ≥ 2).