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Figure 6


Fig. 6. The GSK-3ß–APC pathway is required for integrin-induced regenerative axon assembly from PCL neurons. (A) Overexpression of EGFP had no effect on integrin-mediated axon assembly. (B) Expression of the constitutively activated GSK-3ß mutant (yellow) significantly blocked axon assembly. (C) Axon assembly was abolished in neurons expressing C-EB1 (yellow), a mutant protein that interferes interactions between APC and microtubule plus ends. (D) Axon assembly in neurons expressing C-EB1{Delta}APC was not affected. Arrows indicate transfected cells that express GFP. Axons of transfected neurons are yellow. Neurofilament staining (red) reveals axons of untransfected cells. Magnification, 4x. (E,F) Quantification of axon assembly defect in neurons expressing the active GSK-3ß mutant or the EB1 mutant (*P<0.001 vs control). (G) Laminin stimulation alone was sufficient to phosphorylate GSK-3ß in adult PCL neurons, but had no effect on phospho-ERK and phospho-Akt levels. By contrast, laminin stimulation was unable to phosphorylate GSK-3ß in embryonic day 14 (E-14) DRG neurons in the absence of NGF.





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