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. 2020 Jan 6;9:e52990. doi: 10.7554/eLife.52990

Figure 5. Validation of de novo proteomic comparison of trained vs non-trained.

RC3xCamk2a-Cre mice using BONCAT western blotting BONCAT-WB confirms that in trained mice, there is increased hippocampal synthesis of α- α-adaptin (Ap2a1), neuron specific enolase (NSE: Eno2), V-ATPase subunit B2 (V-ATPase B2: Atp6v1b2), and the α isoform of the structural subunit of protein phosphatase 2A (PP2A-A: Ppp2r1a) compared to non-trained controls. Synthesis of the ARE binding protein ARE/poly(U)-binding/degradation factor 1 (AUF-1: Hnrnpd) was also confirmed to be decreased during spatial LTM formation. The synthesis of the housekeeping gene, glyceraldehyde 3-phosphate dehydrogenase (GAPDH), was unchanged in the hippocampal neurons of trained RC3xCamk2a-Cre mice compared to non-trained controls (p=0.48), together validating the SWATH analysis (n = 4 mice, Student’s paired t-test, *p≤0.05, **p≤0.01, ***p≤0.001).

Figure 5.

Figure 5—figure supplement 1. Complete images of the western blots shown in Figure 5.

Figure 5—figure supplement 1.