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. 2020 May 2;30(12):970–980. doi: 10.1093/glycob/cwaa030

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© The Author(s) 2020. Published by Oxford University Press.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Fig. 1 — Probing substrate glycans of FUT2, FUT6, FUT7 and FUT9 on fetuin (Fet) and asialofetuin (aFet). (A) Strategies for probing substrate glycans of indicated FUTs. FUT2 recognizes both β3- and β4-linked terminal Gal. FUT9 recognizes terminal lactosamine (LN). FUT7 only recognizes sialyllactosamine (sLN). FUT6 recognizes both LN and sLN. Monosaccharide symbols follow the Symbol Nomenclature for Glycans (SNFG) system (PMID 26543186, Glycobiology 25: 1323–1324, 2015) details at NCBI. (B) Probing substrate glycans of FUT2(F2), FUT6(F6), FUT7(F7) and FUT9 (F9) on Fet or aFet with GDP-Cy5-fucose. FUT9 labeled itself (indicated by an arrow). (C) Probing substrate glycans of FUT2, FUT6, FUT7 and FUT9 on Fet or aFet with Alexa Fluor®555 (555), Alexa Fluor®488 (488) and Cy5-conjugated fucose. All reactions in (B) and (C) were incubated at 37^oC for 30 min and then separated on SDS–PAGE and imaged with silver staining (upper panels) and fluorescent imager (lower panels). M, molecular marker.