Figure 3

Splicing variation of NOTCH2. A. Schematic representation of recombinant constructs for full-length (FL) and truncated (TR) NOTCH2 protein isoforms fused with C-terminal Halo-tag protein (marked as Halo). Intracellular part of FL-NOTCH2 consists of two LNRs (Lin-12/Notch repeat domains, marked as N), two NOTCH protein domains (marked as NOD), two ankyrin repeats (marked as ANK) and a topoisomerase II-associated protein domain (marked as PAT1). B. Western blot showing expression of recombinant FL-NOTCH2-Halo and TR-NOTCH2-Halo constructs in HeLa and 293T cell lines. The lanes show two different size markers, negative control (no DNA), mock control (vector only) and results of transfections with FL-NOTCH2-Halo and TR-NOTCH2-Halo in HeLa and 293T cells. C. Confocal imaging of protein expression for the recombinant construct FL-NOTCH2-Halo (red) in HeLa cells. Untransfected cells in the same field serve as negative controls. Antibodies against α-tubulin were used to stain the cytoskeleton (green) while DAPI was used to stain the cell nuclei (blue). D. Confocal imaging of protein expression for the recombinant construct TR-NOTCH2-Halo (red) in HeLa cells. Untransfected cells in the same field serve as negative controls. Antibodies against α-tubulin were used to stain the cytoskeleton (green) while DAPI was used to stain the cell nuclei (blue).