Figure 5.

Analysis of effect of neuraminidase pretreatment on LDL uptake. HepG2 WT cells (A) and CHO WT cells (B) were pretreated with and without neuraminidase and subjected to lectin immunocytology (upper panels) and FITC-LDL binding assays (lower panels). PNA and Maackia amurensis lectin II (MAL II) lectins were used to demonstrate the loss of sialic acids by the neuraminidase treatment. FACS analysis of FITC-LDL binding was performed with HepG2 and CHO WT and SC cells, and pretreatment with neuraminidase affected binding to WT cells but not to SC cells, which is in agreement with the established O-glycosylation capacity of these cells where WT produces sialylated core 1 O-glycans, and SC produces the nonsialylated GalNAc O-glycans. The experiment was performed twice with similar results.