Figure 2
Activation of human NOTCH1 by transient co-transfection of TM2D3. (a) Activation of human NOTCH1 by transient co-transfection of a TM2D3v2 construct in 293 T cells. Experiments were conducted as described in Fig. 1a. (b) Statistical analysis of the activation of human NOTCH1. Two rounds of independent experiments were additionally conducted as is shown in a. The adjusted volume of the NICD band in each extract was normalized to that for vinculin in the corresponding lane. In each experiment, the values in the sample transfected with the vector for NOTCH1 and in empty vector control for TM2D3 were set as 1. The values for the other samples were calculated relative to these amounts. The error bars indicate the standard deviations (n = 3). F(2,6) = 8.3; P = 1.9 × 10–2 using a one-way ANOVA. * P = 1.3 × 10–2 by Dunnett’s multiple comparison test. (c) Activation of a NOTCH1 construct by co-transfection of TM2D3 in U2OS cells. Cells were transiently transfected with expression vectors for the indicated proteins, a plasmid containing a reporter, and a plasmid containing an internal control. Activation of NOTCH1 receptor was assessed by a luciferase assay. The error bars indicate the standard deviations (n = 3). *P = 6.9 × 10–5; **P = 6.7 × 10–3 by Dunnett’s multiple comparison test. The NOTCH1 construct, the reporter, and the internal control are indicated schematically below the graph.
