Figure 1
From: Inhibition of Wnt signalling by Notch via two distinct mechanisms
Characterization of ΔEGF_N1 and NICD constructs. (A) Schematic showing the structure of the full length, ΔEGF_N1 and NICD proteins. (B) Western blot analysis of ΔEGF_N1, NICD constructs and empty vector. Expressed proteins were detected by probing the western blot with an antibody that recognises the myc epitope tag found within both proteins. Renilla luciferase is shown as a loading control. The position of molecular weight markers is shown in KDa. (C) Immunofluorescence analysis of ΔEGF_N1 and NICD proteins. Scale bar is 25 μm. (D) Unlike NICD, ΔEGF_N1 failed to activate Notch signalling. HEK293T cells were transfected with p10xRbpj-luc either alone or with plasmids encoding the ΔEGF_N1 and NICD proteins. Experiments were performed in triplicate. pRL-CMV was used as a transfection control and cells were lysed 48 h post transfection to determine luciferase activity. Data are presented as mean fold change (± SEM) in RLU (NS P > 0.05; ****P < 0.0001 one-way ANOVA and Tukey’s post-hoc test, N = 3). Original uncropped Western Blot images of panel B are shown in Supplementary Fig. 5
