Fig. 5.
NOTCH3L1519P produces ICD in a ligand-independent manner. (A) Western blot analysis of FL, TMIC and NEXT and ICD fragments from cell extracts using an anti-NOTCH3 ICD antibody, following activation of the cells by immobilized jagged2 (Jag2-Fc) and treatment with the γ-secretase inhibitor DAPT or MG132, as indicated. (B) NIH3T3 cells were transfected with wild-type, NOTCH3L1519P or control plasmid together with 12XCSL-luc reporter and β-gal plasmids and cultured on immobilized jagged2 (Jag2) in combination with treatment by DMSO or DAPT, as indicated (n=3). Statistical analysis was performed from three experiments using paired Student's t-test, *P<0.1, **P<10−2, ***P<10−3, RLU, relative luminescence units.