Figure 1.

Intra- and interday repeatability of the optimized method. (A) Workflow of the optimized method. A scheme detailing the chemistry used for the O-glycan release, hydrazide enrichment, and 2-AB labeling can be found in Supporting Figure S5. (B) The O-glycans from approximately 2.5 × 10⁵ HaCaT^WT cells were released, labeled, and analyzed by C18 LC–MS/MS for two times with six technical replicates on two successive days. Displayed are average relative intensities for the glycans with a relative abundance above 1% per day, with error bars representing the standard deviations. Graphics in panel (A) were created using https://biorender.com/. H, hexose; N, N-acetylhexosamine; F, fucose; S, N-acetylneuraminic acid.