Figure 3

Characterization of signals in Glycosyl transferase 25 domain 1, suggesting ER residence. A) Summary of GLT25D1 expression constructs. HA indicates Hemagglutinin epitope tag; Myc refers to Myc epitope tag; FL denotes a full-length protein; RDEL represents carboxy-terminal Arg-Asp-Glu-Leu sequence; SS stands for signal sequence and Δ is followed by deleted part of the protein. B) Co-transfected Huh7 cells with constructs GLT25D1 FL-HA and GLT25D1 ΔSS-Myc were subjected to immunofluorescence analysis. Signal of FL-HA is shown in green and of ΔSS-Myc in red. C) The constructs 1 to 9 shown in A were transfected into Huh7 cells. The total cell lysates (Total) and the culture medium (Medium) from these cells were analyzed by SDS-PAGE and western blot. Using anti-bodies against the N- and C-terminal tags of the constructs, anti-HA and anti-Myc, the proteins were detected.