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. 2008 Mar 10;180(5):915–929. doi: 10.1083/jcb.200707126

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Copyright © 2008, The Rockefeller University Press

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Figure 6. — SIRT2 overexpression impairs ephrin-A–mediated growth cone collapse. (A) Hippocampal neurons were coelectroporated with constructs expressing the indicated SIRT2 proteins and a GFP vector. The cells were cultivated on laminin for 2 d and then treated with 1 μg/ml ephrin-A5-Fc or Fc for 30 min and subsequently stained for F-actin. (B) The percentage of collapsed growth cones was quantified by morphological criteria. (C) Summary of the overall change in the growth cone collapse rate. The percentage of growth cone collapse in response to ephrin-A5-Fc was subtracted from the response to Fc alone. (D) Neurons were coelectroporated with constructs expressing the indicated proteins. The cells were then stained for F-actin and the fluorescence intensity of individual growth cones was determined. Untreated growth cones contain high F-actin levels, which are decreased upon ephrin-A5 treatment. The mean values and SD of 50 or more individual growth cones/conditions are displayed. (E) Neurons were coelectroporated with constructs expressing siSIRT2 or siLUC. Growth cones were stained for β-tubulin (left, red) and F-actin (left, green). Quantification of the mean number of filopodia/growth cones after siSIRT2 or siLUC treatment is shown on the right. (F) Model of SIRT2 function and regulation by Cdk complexes. For details, see Discussion. Error bars represent SD. **, P < 0.01; ***, P < 0.001.