Figure 4
From: Fluorescent glycan fingerprinting of SARS2 spike proteins
Fingerprinting by ST3Gal6 and ST6Gal1 on various SARS2 Spike proteins. Glycans of various recombinant SARS2 spike proteins released by PNGase F with (+) or without (−) neuraminidase pretreatment were labeled by ST3Gal6 or ST6Gal1 together with FUT8 and separated on a 17% SDS gel. The gel was then imaged with silver staining (upper panel) or fluorescent imaging (middle and lower panel in different contrasts). Bands in red are complex or hybrid glycans labeled by ST6Gal1 or ST3Gal6 (via the incorporation of Cy5-Neu5Ac). Bands in green are oligo-mannose glycans labeled by FUT8 (via the incorporation of Alexa Fluor 555-conjugated Fucose). For oligo-mannose labeling, MGAT1 and UDP-GlcNAc were also included in the labeling mix to convert the glycans to the substrates for FUT8. RC, RBD domain of the SARS2 Spike protein expressed in CHO cells; RH, RBD domain of the SARS2 Spike protein expressed in HEK293 cells; SH, whole SARS2 Spike protein expressed in HEK293 cells.
