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| | Immunofluorescense analysis, transfected COS-7 cells |
| | Cells were transfected on cover-slips with the appropriate MAN2B1-construct and 36-42 h post transfection protein synthesis was stopped by incubation with cycloheximide for 2 h followed by fixation in ice-cold methanol. MAN2B1 was detected using using antiserum against denatured bovine liver MAN2B1 and visualized using FITC-conjugated secondary antibody. LAMP-1 was used as lysosomal marker and detected by antibody H4A3; PDI was used as ER-marker and detected by anti-PDI antibodies, both markers were visualized using TRITC-conjugated secondary antibody. |
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