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. 2020 Apr 9;21(7):2609. doi: 10.3390/ijms21072609

Figure 6.

Figure 6

The loss of function/missense mutation of Ebp1 gene decreases GAD67 expression and disturbs epigenetic control. (A) Schematic presentation of two of loss of function mutations, mutation to stop codon, from SZ patients is shown. (B) RNA and protein expression levels of Gad67, Hdac1 and Dnmt1 were measured in transfected Ebp1(−/−) MEF cells. GFP-mock, Ebp1 WT, or Ebp1 G183Ter constructs were used for transfections. The RNA expression was tested using qRT-PCR with specific primer sets. The protein level was measured using IHC with anti-GAD67, HDAC1. and DNMT1 antibodies. (C) Ebp1(−/−) MEF cells were co-transfected with pGL-Gad67 promoter along with control or Ebp1 WT, G183Ter mutation construct plasmids. Luciferase activity of GAD67 was measured using the luciferase assay. ** p < 0.001. (D) Luciferase activity of Hdac1 (left) and Dnmt1 (right) were measured in the transfected Ebp1(-/-) MEF cells which over-expressed Ebp1 WT or G183Ter mutation proteins. ** p < 0.001. (E) Schematic illustration shows molecular mechanisms of EBP1 in GAD67 epigenetic effects. Created with BioRender. ** p < 0.001. All data are shown as the mean ± s.e.m.