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Fig. 2. GSK-3ß phosphorylated at Ser9 does not accumulate at the tip of neurites in stage-2 neurons or in the axon of stage-3 neurons. (A) Confocal single planes of an individual neuron transfected with GFP and stained for phosphorylated Ser9 GSK-3ß. The ratio image of the two is shown in the right picture. Bar, 10 µm. (B) Quantification of the results of A. Hippocampal neurons were fixed at different stages of development and total GSK-3ß or GSK-3ß phosphorylated at Ser9 visualised using specific primary and fluorescent secondary antibodies. The fluorescence signal was divided by the signal obtained from the soluble cytoplasmic marker GFP or the DTAF signal, and the ratio of intensities in the tips was determined, normalised and sorted according to intensity. As control the fluorescent intensity of a soluble protein that does not accumulate in individual tips (p38) was used (mean ± s.e.m., *P<0.005).
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