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On DIV12 the neurons were fixed (4% paraformaldehyde, 3% sucrose, 60 raM PIPES, 25 mM HEPES, 5 mM EGTA, 1 niM MgCl 2 , pH 7.4) for 20 min at room temperature and mounted

Proteins (1030 g/well, equal amounts within group but different amounts among groups for better comparison) were separated on 10% or 12% SDS-PAGE gel and transferred on an equilibrated polyvinylidene difluoride membrane (PVDF, Amersham Biosciences, Buckinghamshire, UK)

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