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. Author manuscript; available in PMC: 2007 Aug 13.
Published in final edited form as: J Neurochem. 2001 Sep;78(6):1219–1232. doi: 10.1046/j.1471-4159.2001.00495.x

Fig. 6.

Fig. 6

Growth factor stimulation of CREB DNA binding activity is blocked at multiple time points in SH-SY5Y cells overexpressing GSK3β. (a) CREB DNA binding activity was measured in nuclear extracts prepared from wild-type, vector-transfected, and HAGSK3β-overexpressing SH-SY5Y cells incubated without serum (24 h) and treated without or with EGF (50 ng/mL) or IGF-1 (50 ng/ mL). Quantitative values are expressed as the percent of basal CREB DNA binding activity in serum-starved cells. Mean ± SEM, n = 3–4. (b) GSK3β activity was measured by immunoprecipitating GSK3β, incubating the immunoprecipitated GSK3β with recombinant human tau and [32P]ATP, and measuring the phosphorylation of tau, as described in Materials and methods, in wild-type (wt) cells, or in HA-GSK3β-overexpressing cells (HA-GSK3β) incubated without serum (24 h) and treated without (basal) or with EGF (50 ng/mL; 1 h) or IGF-1 (50 ng/mL; 1 h).