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(A) Schematic depiction of a glycosylated membrane protein. (B) The <t>lectin-based</t> approach relies on the five lectins WGA, SNA, PHAL, AAL and <t>PSA,</t> <t>conjugated</t> to the DNA-PAINT docking sequences R1-R5. (C) The metabolic labeling-based approach relies on metabolic incorporation of N-azidoacetylmannosamine, which enables stochastic labeling of sialic acids with a mixture of the DNA-PAINT docking sequences R1-R5. (D) Protein (blue) and lectin localizations (red/green) are resolved at conventional DNA-PAINT resolution for the lectin-based approach. (E) Protein localizations (blue) are resolved at DNA-PAINT resolution and sugar residues (purple) are resolved at RESI resolution for the metabolic labeling-based approach. Schematics are not to scale. Scale bars in (D) and (E): 15 nm.
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(A) Schematic depiction of a glycosylated membrane protein. (B) The lectin-based approach relies on the five lectins WGA, SNA, PHAL, AAL and PSA, conjugated to the DNA-PAINT docking sequences R1-R5. (C) The metabolic labeling-based approach relies on metabolic incorporation of N-azidoacetylmannosamine, which enables stochastic labeling of sialic acids with a mixture of the DNA-PAINT docking sequences R1-R5. (D) Protein (blue) and lectin localizations (red/green) are resolved at conventional DNA-PAINT resolution for the lectin-based approach. (E) Protein localizations (blue) are resolved at DNA-PAINT resolution and sugar residues (purple) are resolved at RESI resolution for the metabolic labeling-based approach. Schematics are not to scale. Scale bars in (D) and (E): 15 nm.

Journal: bioRxiv

Article Title: Spatial mapping of cell-surface protein glycosylation at molecular resolution

doi: 10.1101/2025.09.30.679542

Figure Lengend Snippet: (A) Schematic depiction of a glycosylated membrane protein. (B) The lectin-based approach relies on the five lectins WGA, SNA, PHAL, AAL and PSA, conjugated to the DNA-PAINT docking sequences R1-R5. (C) The metabolic labeling-based approach relies on metabolic incorporation of N-azidoacetylmannosamine, which enables stochastic labeling of sialic acids with a mixture of the DNA-PAINT docking sequences R1-R5. (D) Protein (blue) and lectin localizations (red/green) are resolved at conventional DNA-PAINT resolution for the lectin-based approach. (E) Protein localizations (blue) are resolved at DNA-PAINT resolution and sugar residues (purple) are resolved at RESI resolution for the metabolic labeling-based approach. Schematics are not to scale. Scale bars in (D) and (E): 15 nm.

Article Snippet: A lectin cocktail (2.5 μg/mL of each lectin, conjugated to DNA strands R1 to R5 by Massive Photonics) prepared in 1X tris buffer (Fisher Bioreagents, ref# M-15836) was applied to the cells at room temperature for 30 min, allowing specific binding to glycan targets on the cellular surface.

Techniques: Membrane, Labeling