Fig. 2: GluN2A-dependent NMDAR signaling regulates nuclear DNMT3A1 protein levels in 14–15 DIV hippocampal primary neurons. | Neuropsychopharmacology

Fig. 2: GluN2A-dependent NMDAR signaling regulates nuclear DNMT3A1 protein levels in 14–15 DIV hippocampal primary neurons.

From: Synaptic control of DNA methylation involves activity-dependent degradation of DNMT3A1 in the nucleus

Fig. 2

a, b Treatment with the NMDAR antagonist APV (20 μM) prevented the reduction in the nuclear levels of DNMT3A1 following synaptic stimulation by Bic/4AP for 6 h. c, d Treatment with the GluN2A inhibitor NVP-AAM077 (50 nM) prevented the reduction in the nuclear levels of DNMT3A1 following synaptic stimulation by Bic/4AP for 6 h. Two-way ANOVA followed by Bonferroni’s post hoc test. e Timeline view of hippocampal neuron treatment by Bic/4AP for 6 h with or without NVP-AAM077. f, g Quantitative immunoblotting revealed the inhibition of DNMT3A1 degradation by the use of NVP-AAM077. One-sample t-test is performed while the hypothetical value is set to 100. h, i shRNA-based knockdown of GluN2A in the presence of synaptic stimulation by Bic/4AP for 6 h prevented the DNMT3A1 degradation. j, k Treatment with the GluN2B subunit inhibitor ifenprodil (10 μM) in the presence of synaptic stimulation with Bic/4AP for 6 h did not prevent the reduction in the nuclear levels of Dnmt3A1. Scale bars, 20 μm. Two-way ANOVA followed by Bonferroni’s post hoc test. ***p < 0.001, n.s. not significant. Error bars present S.E.M. Sample numbers for each experimental group indicate neurons from three different culture preparations.

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