Heparan sulfate (HS) is a polysaccharide fundamentally important for biologically activities. T/Tn antigens are universal carbohydrate cancer markers. Here, we report the specific imaging of these carbohydrates using a mesenchymal stem cell line and human umbilical vein endothelial cells (HUVEC). The staining specificities were demonstrated by comparing imaging of different glycans and validated by either removal of target glycans, which results in loss of signal, or installation of target glycans, which results in gain of signal. As controls, representative key glycans including O-GlcNAc, lactosaminyl glycans and hyaluronan were also imaged. HS staining revealed novel architectural features of the extracellular matrix (ECM) of HUVEC cells. Results from T/Tn antigen staining suggest that O-GalNAcylation is a rate-limiting step for O-glycan synthesis. Overall, these highly specific approaches for HS and T/Tn antigen imaging should greatly facilitate the detection and functional characterization of these biologically important glycans.

中文翻译：

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通过点击化学使用糖基转移酶对特定的细胞聚糖结构进行成像

硫酸乙酰肝素（HS）是一种对生物学活性至关重要的多糖。T / Tn抗原是通用的碳水化合物癌症标志物。在这里，我们报告使用间充质干细胞系和人脐静脉内皮细胞（HUVEC）的这些碳水化合物的具体成像。通过比较不同聚糖的成像证明了染色特异性，并通过去除目标聚糖（导致信号丢失）或安装目标聚糖（导致信号增益）进行了验证。作为对照，还成像了代表性的关键聚糖，包括O-GlcNAc，乳糖胺基聚糖和透明质酸。HS染色揭示了HUVEC细胞的细胞外基质（ECM）的新型结构特征。T / Tn抗原染色的结果表明，O-GalNAcylation是O-聚糖合成的限速步骤。总体而言，这些用于HS和T / Tn抗原成像的高度特异性方法应极大地促进这些生物学上重要的聚糖的检测和功能表征。