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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 5. — SIRT2 modulates neurite outgrowth of hippocampal neurons. (A) Hippocampal neurons were coelectroporated with constructs expressing the indicated SIRT2 proteins and a GFP vector and cultivated for 2 d in vitro, followed by staining for acetylated α-tubulin. Arrowheads identify some of the GFP-expressing cells. Bars, 50 μm. (B) Quantification of experiments exemplified in A. The neurite length was determined in response to the expressed proteins indicated. The neurite length of neurons electroporated with the parental vector was set to 100%. Only neurons with measurable neurites were taken into account. Relative neurite length is expressed in percentage of control (set to 100%). (C) Neurons without any detectable neurite growth were determined in response to the indicated SIRT2 proteins. (D) The level of acetyl-α-tubulin in SIRT2-S331A–expressing and control electroporated neurons was compared. A total of 60 neurons (20 in each of three independent experiments) were captured and acetylated α-tubulin levels measured using AxioVision software. The values from control cells were set to 100%. (E) Neurite length was measured, as in A, in response to coexpressed p35–Cdk5. (F) Neurons were coelectroporated with vectors expressing GFP and the indicated siRNAs. The neurite length of neurons electroporated with siLUC was set to 100%. Relative neurite length is expressed in percentage of control (set to 100%). (G) The neurites were classified into four length groups, from very short (0–50 μm) to very long (>150 μm). The percentage of neurons within each group is displayed. Error bars represent SD. **, P < 0.01; ***, P < 0.001.