Skip to main content
. 2020 Jul 29;45(12):2120–2130. doi: 10.1038/s41386-020-0780-2

Fig. 1. Synaptic activity regulates DNMT3A1 protein levels in neurons.

Fig. 1

a Nuclear DNMT3A1 immunofluorescence is prominent in MAP2-positive neurons but much less in GFAP-positive astrocytes at 15 DIV hippocampal cultures. Scale bars are 20 μm. b, c Downregulation of DNMT3A1 protein levels in 14–15 DIV hippocampal primary neurons following treatment with Bic/4AP for 1 h, 3 h or 6 h as evidenced by quantitative immunocytochemistry. Scale bar is 20 μm. Unpaired Student’s t-test **p < 0.01, ***p < 0.001. d–f DNMT3a1 protein levels are decreased following the removal of tonic inhibition in DIV 21 cortical neurons. β-Actin was used as an internal control for normalization. Student’s t-test *p < 0.05. g, h Hippocampal neurons were treated with Bic/4AP for 10 min or 3 h. Media from 10 min-long treated neurons was washed out and cells were kept for 3 h before fixation. Both treatments were equally effective to reduce nuclear DNMT3A1 immunofluorescence. Application of 100 μM NMDA for 10 min had no effect. Unpaired Student’s t-test **p < 0.01, ***p < 0.001. Error bars present S.E.M. Sample numbers for each experimental group indicate neurons from three different culture preparations.