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Fig. 6. Krm increases cell-surface levels of LRP6. (A) Western blot
analysis of HEK293T cells transfected with flag-LRP6 and the
indicated constructs. Samples were treated with EndoH as indicated. Arrows
indicate the EndoH-resistant mature and EndoH-sensitive immature forms of
LRP6. (B) Cell surface biotinylation assay. HEK293T cells were
co-transfected with flag-LRP6 and the indicated constructs. After
cell surface biotinylation, cell lysates were immunoprecipitated with
anti-flag antibody and subjected to SDS-PAGE and western blot analysis.
Membranes were probed with streptavidin-HRP to detect plasma membrane LRP6
(upper panel) and anti-flag antibody to detect total LRP6 (lower panel).
(B') The cytoplasmic protein NME1 is not biotinylated (upper
panel). Lower panel: Total NME1 protein. dg, deglycosylated form; im,
EndoH-sensitive immature form; ma, EndoH-resistant mature form.