A Glycosphingolipid Binding Domain Controls Trafficking and Activity of the Mammalian Notch Ligand Delta-Like 1
Figure 6
Dll1AG does not recycle and exhibits a shorter half-life than wild type Dll1.
(A) Surface proteins of Dll1- and Dll1AG-expressing OP9 cells were labeled with biotin and a recycling assay was performed as described in Materials and Methods. NDB: not debiotinylated; Mesna2: second Mesna treatment. (B) Western blot analysis of cells expressing Dll1 or Dll1AG. Approximate equal loading of the full-length form was used in order to facilitate estimation of the extent of metalloprotease cleavage. (C) HeLa cells were transiently-transfected with plasmids encoding Dll1-Apa or Dll1-Apa-AG together with GFP. Following cycloheximide treatment the levels of ligands were monitored as described in Figure 2. Western blots were performed using anti-Dll1 and anti-GFP antibodies.