Figure 3

Evaluation of the proteins generated containing the C-terminal half of RAI1. A) Schematic representation of the construction of proteins 1038-end and 1229-end that were generated by PCR. Both proteins are tagged with the HA peptide on the C-terminal end. In blue is represented the Poly-Gln domain, in yellow: Poly-Ser domains, in black: in silico described nuclear localization signals, in slanted line: the PHD domain. B) The molecular weight for both proteins was obtained by transfecting them in Neuro-2a cells and then a western blot analysis was performed with anti HA antibody. Molecular weight for the proteins 1038-end and 1229-end are depicted and also are shown the controls with only the transfection of empty vector (e/v) and untransfected Neuro-2a cells (u/t). C) The graphic represents the percentage of activation for 1038-end protein (grey) and 1229-end protein (white) compared to the wild type full-length protein (black). Values represent mean +/- SEM. (* depicts statistically significant differences, p ≤ 0.0002). D) Each plasmid was transfected in Neuro-2a cells and an immunofluorescence was performed with anti HA antibody. Nuclei staining were made with DAPI. The table shows subcellular localization for 200 cells positive for anti HA. α = antibody against HA.