Supplemental Material

This article contains the following supporting material:

• Supplemental Figure 1 - Levels of Nb1 over-expression or downregulation correlate with the severity of the observed phenotype
  Dissociated hippocampal neurones were transfected prior to plating, fixed after 1 or 3 days in culture and stained for GFP (green), F-actin (red) and tubulin (TUJ1, blue). (A) After 1 day in culture, neurones overexpressing high levels of Nb1 (arrow) exhibited a simplified morphology compared to neurones expressing moderate amounts of Nb1 (arrowhead). (B) Neurones expressing high levels of Nb1 shRNA (arrow), and therefore low levels of endogenous Nb1, typically exhibited splayed growth cones whereas neurones expressing low levels of shRNA (arrowhead) appeared less affected. (C) After 3 days in culture, neurones expressing the highest levels of Nb1 shRNA (arrow) were more affected than neurones expressing low levels of the constructs (arrowhead). (D) and (E) are magnifications of (C) illustrating the morphological differences between neurones expressing low (D) and high (E) levels of Nb1 shRNA. Size bars (A, C): 45 µm, (B): 25 µm, (D, E): 30 µm.