Additional file 3 of Ubiquitylation is required for the incorporation of the Notch receptor into intraluminal vesicles to prevent prolonged and ligand-independent activation of the pathway

Additional file 3: Figure S3. N-HA and NK2R-HA ubiquitination assay. (A, B) Ubiquitination-assay of N-HA and NK2R-HA. The variants were co-expressed in S2 cells with Flag tagged Ubiquitin (Flag-Ubi) and Su(dx) (A) or Dx (B) (n= 3). Full length N-HA and NK2R-HA bands run at 300 kDa, that of the activated intracellular domains at 110 kDa. The results show that N-HA is ubiquitinated by Su(dx) (upper left panel), as well as Dx (lower left panel). In contrast, NK2R-HA was not ubiquitinated by Su(dx) (upper left panel) or Dx (lower left panel). Right panels show whole cell extracts. Peanut antibody staining was used as loading control. All samples used for the IP showed a similar protein level.