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| | Confocal immunofluorescence analysis, transfected HeLa cells |
| | Cells were grown on cover-slips and transfected with the appropriate MAN2B1-construct. 36-42 hrs post transfection, protein synthesis was stopped by incubation with cycloheximide for 2 hrs followed by fixation in ice-cold methanol. MAN2B1 was detected using antiserum against denatured bovine liver MAN2B1 and visualized using Alexa568-conjugated secondary antibodies. 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 Alexa488-conjugated secondary antibodies. |
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