Fig. 3. Downregulation of SIRT2 increases the levels of Ehmt2 and H3K9me2 at the promoters of presynaptic genes in hippocampal neurons. (A) Representative blots of SIRT2 and CoREST following immunoprecipitation of primary hippocampal neuron extracts (left). Protein levels were normalized to the abundance in SIRT2 input (right). Reduced SIRT2 binding to CoREST in lenti-shSIRT2 infected neurons (n=4). (B) ChIP assays. SIRT2 knockdown decreased the association of SIRT2-CoREST complexes with the Ehmt2 and Scmh1 promoters in hippocampal primary neurons (n=3). (C, D) Levels of Ehmt2 and Scmh1 mRNAs in mouse hippocampal primary neurons were assayed by qPCR (n=3). (E) ChIP assays. SIRT2 knockdown increased H3K9me2 levels of Egr1, Synapsin1 (Syn1) and Synaptophysin (SYP) promoters in hippocampal primary neurons (n=3). (F, G) Synapsin1 and Synaptophysin mRNAs in the mouse hippocampal primary neurons were assayed by qPCR (n=3). (H) Effect of AGK2 treatment on levels of Ehmt2 and Scmh1 mRNAs in mouse hippocampal primary neurons was assayed by qPCR (n=3). (I, J) Effect of AGK2 treatment on levels of Synapsin1 and Synaptophysin mRNAs in mouse hippocampal primary neurons was assayed by qPCR (n=3). Data are mean±s.e.m. Unpaired t tests; *p<0.05, **p<0.01, ***p<0.001.
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