Figure 4
Molecular evaluation of two point mutations associated with SMS. A) Schematic representation RAI1 Q1562R and RAI1 S1808N. 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. The coding sequence for HA epitope is represented in red. B) The molecular weight of mutated proteins was calculated in Neuro-2a cells by western blotting with anti RAI1 antibody. The obtained molecular weight is depicted and also the controls for the immunoreactivity are shown (e/v: extracts transfected only with the empty vector and u/t represents untransfected cells control). C) The percentage of the reporter transcription activation is shown for RAI1-HA Q1562R (white) and RAI1-HA S1808N (grey) compared to RAI1-HA wild type protein (black). Values represent mean +/- SEM. D) Each plasmid was transfected in Neuro-2a cells and immunofluorescence was performed with anti RAI1 antibody and nuclei staining is shown with DAPI. The table represents subcellular localization of 200 cells immunodetected with anti RAI1 antibody. α = antibody against.