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. 2016 May 18;6:26119. doi: 10.1038/srep26119

Figure 4. Insulin stimulation enhances GluR1 phosphorylation by forskolin.

Figure 4

(A–C) Immunoblots of hippocampal slices from APP, ApoE3xAPP (E3XAPP) and ApoE4xAPP (E4XAPP) mice were treated with 50 μM of forskolin (FSK) with or without pre-treatment with 1 μM of insulin. Each blot is a representative of six independent experiments. Blot images were cropped for comparison and all relevant gels have been run under similar experimental conditions. Cropping area is indicated by black line surrounding the border of the blot figures. Results were expressed as a fold change in protein expression as compared to the non-treated slices within the same mouse line. Error bars represent ± SEM (n = 6 slices). (A) At 26 weeks, FSK treatment significantly increased P-GluR1 S831 (**p < 0.01) and P-GluR1 S845 (**p < 0.01) expression in ApoE3xAPP and APP mice. Insulin pre-treatment further potentiated P-GluR1 S831 and P-GluR1 S845 expression in ApoE3xAPP and APP mice when compared to non-treated slices (**p < 0.01). This insulin stimulated increase was significantly higher than FSK treatment alone (#p < 0.05, ##p < 0.01). (B) At 52 weeks, only slices from ApoE3xAPP mice were responsive to FSK treatment. When compared to non-treated slices within the same mouse line, FSK treatment significantly increased P-GluR1 S831 and S845 expression (*p < 0.05). Insulin pre-treatment further potentiated P-GluR1 phosphorylation in ApoE3xAPP mice (**p < 0.01) as compared to non-treated slices. When compared against slices that were treated with only FSK, insulin stimulation led to a greater increase in P-GluR1 expression in ApoE3xAPP mice (#p < 0.05, ##p < 0.01). (C) At 78 weeks, FSK treatment significantly increased GluR1 phosphorylation at S831 and S845 in ApoE3xAPP mice (*p < 0.05). As compared to non-treated slices, insulin pre-treatment potentiated FSK-induced GluR1 phosphorylation in ApoE3xAPP mice (*p < 0.05, **p < 0.01). When compared against slices that were treated with FSK only, insulin stimulation only led to a greater increase in phosphorylated GluR1 S831 expression in ApoE3xAPP mice (#p < 0.05).