Fig. 4: Ischemic stroke results in ubiquitination of postsynaptic kinases and phosphatases.

a Proteins with increased and decreased post-ischemic ubiquitination were assessed for GO enrichment for biological process. Benjamini-corrected P values for the top four terms for the “Ub increased” dataset are depicted. b All proteins with increased ubiquitination after ischemia and enrichment in the four top categories for biological process were investigated for GO enrichment for molecular function and cellular components. Benjamini-corrected P values for the five highest-ranking terms are shown. c Post-ischemic ubiquitination of select PSD-associated kinases and phosphatases was confirmed by immunoprecipitation of proteins of interest from ipsi- and contralateral detergent-insoluble cortical lysates from MCAO/1 h reperfusion-treated mice and detection of ubiquitin by Western Blotting. IgG-isotype antibodies served as controls. Results from n = 3 mice/group were quantified. CaMKIIα: *P = 0.0009; CaMKIIβ: *P = 0.0040; PKCβ: *P = 0.0021; PKCγ: *P = 0.0099; Cdk5: *P = 0.0015; Pyk2: *P = 0.0016; CKβ: *P = 0.0002; Pten: *P = 0.0013. Two-tailed unpaired t-test. Data are expressed as mean ± s.e.m. d Domain structure of prominent PSD-associated kinases and phosphatases found ubiquitinated after ischemia. Numbers represent amino acid positions, and arrows indicate ubiquitinated residues identified by MS analysis. BP biological process, c contralateral, C carboxy-terminus, C1a, and C1b diacylglycerol-binding domain, C2 calcium-binding domain, CaM calmodulin, FAT focal adhesion kinase-targeting domain, FERM, 4.1 protein, Ezrin radixin, and moesin domain, GO gene ontology, i ipsilateral, LTP long-term potentiation, N amino-terminus, nc not called, P proline-rich region, PDZ-b PSD95, Dlg1 Zo-1-containing domain-binding domain, POI protein of interest, Ub ubiquitination.