Figure 4
Notch proteins are indispensable to DSL ligand-induced increase in synaptic vesicle proteins.
(a) Immunoblot analysis of neuronal culture from NTKO (Notch1flox/flox; Notch2flox/flox; Notch3−/−) mice. Infection of the recombinant adenovirus (AdV) encoding Cre recombinase (Cre) reduced the protein levels of Notch1 and 2 compared to that in cells infected with control adenovirus (LacZ). The gels have been run under the same experimental conditions and representative blots are cropped and used in this figure. Full-length gels are presented in Supplementary Fig. S3. (b) Immunoblot analysis of AdV-infected neurons cocultured with NIH3T3 cells. The increase in the levels of synaptophysin 1 (syp) and VGLUT1 by Jagged1 or Delta-like1 was abolished by the infection of Cre AdV. The gels have been run under the same experimental conditions and representative blots are cropped and used in this figure. (c) Immunoblot analysis of wild type primary neuronal culture infected with Cre AdV were cocultured with NIH3T3 cells. (d, e) Hippocampal neurons from NTKO mice (d) or wild type mice (e) were transfected with myristoylated Venus (green) with or without Cre recombinase and fixed at 10 DIV. The intensities of VGLUT1 immunofluorescence (red) along axon of neurons overexpressing Venus were quantified and shown below the panels (n = 5–10, means ± SEM, *p < 0.05 by student’s t-test). Scale bars, 5 μm.
