Fig. 5: CaMKIIα hyperphosphorylation in the Adnp-HT hippocampus.

a A volcano plot showing Adnp-HT SynGO-PTM-DEPPs (25 proteins found in both PTM-DEPs and SynGO proteins; p < 0.05 + FC > 2.0) plotted by their changes in PTM levels. The protein-encoding gene names are indicated. Note that both CaMKIIα and CaMKIIβ are hyperphosphorylated. b Validation of CaMKIIα hyperphosphorylation by immunoblot analysis. Adnp-HT lysates (crude synaptosomal/P2 fraction; 4 months) were immunoblotted for total CaMKIIα, Thr286-phosphorylated CaMKIIα (p-CaMKIIα), and β-actin (control) (n = 9 [WT], 9 [HT], Welch’s t-test for p-Camk2a and p-Camk2a/Camk2a, Student’s t-test for Camk2a). c Normalization of HFS-LTP at Adnp-HT Schaffer collateral (SC)-CA1 synapses (3–4 months) by the CaMKIIα inhibitor KN-62 (2.5 µM). Note that we lowered KN-62 concentration down to 2.5 µM and limited KN-62 treatment to 20 min before LTP induction to maximize the genotype difference in KN-62-dependent LTP inhibition. Gray, black, red, blue, and green colors in the voltage traces indicate baseline (in WT-DMSO/HT-DMSO/WT-KN-62/HT-KN-62), LTP (WT-DMSO), LTP (HT-DMSO), LTP (WT-KN-62) and LTP (HT-KN-62) synapses, respectively (n = 15 slices from 10 mice [WT-DMSO], 12, 9 [WT-KN-62], 14, 8 [Adnp-HT-DMSO], and 13, 10 [Adnp-HT-KN-62], three-way ANOVA for 60 min and two-way RM-ANOVA with Turkey’s multiple comparison test for last 10 min). Significance is indicated as *(<0.05), **(<0.01), ***(<0.001), or ns (not significant).