Figure 4

N-linked glycosylation of Glycosyl transferase 25 domain 1. A) Full length GLT25D1 (FL) or GLT25D1 with the signal sequence deleted (ΔSS) constructs were transfected into Huh7 cells. Both constructs were carboxy terminal tagged with MycHis6, to purify proteins using Co²⁺-beads and to detect in western blotting. Cell lysates are shown in the first two lanes after separation on SDS-PAGE and western blotting using an antibody against Myc. The purified proteins were subjected to either none (-) or Endoglycosidase H (EndoH) or F (EndoF) treatment (See for more details Methods). The results for FL protein are shown in lanes 3 to 5 and for ΔSS protein in lanes 6 to 8. B) Partial de-glycosylation treatment of GLT25D1 full length (FL).