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. 2021 Oct 29;297(6):101372. doi: 10.1016/j.jbc.2021.101372

Figure 5.

Figure 5

REST interacts with CREB and CBP/p300 proximately to enhance EAAT2 transcription in astrocytes.A–C, REST directly interacts with CREB and CBP/p300 in astrocytes, showed by the proximity ligation assay (PLA) as described in the Experimental procedures section (×40 magnification with confocal microscope, the scale represents 10 μm). Insets show a higher magnification of the PLA puncta in the nuclear region (the scale represents 5 μm). D–G, H4 astrocytes were cotransfected with the EAAT2 promoter vector and REST with CREB (D), CBP/p300 (E), HDAC1 (F), or HDAC4 (G), followed by measurement of EAAT2 promoter activities by luciferase assay. H, H4 astrocytes were overexpressed with REST, followed by coimmunoprecipitation for CBP/p300, p-CREB, HDAC1, and HDAC4 in the nuclear extract. ∗∗p < 0.01; ∗∗∗p < 0.001; ∗∗∗∗p < 0.0001; ####p < 0.0001, compared with the controls; @@@p < 0.001; @@@@p < 0.0001, compared with each other (one-way ANOVA followed by Tukey's post hoc test; n = 3). Data are expressed as mean ± SD. The data shown are representative of three independent experiments. CBP, CREB-binding protein; CREB, cAMP response element–binding protein; EAAT2, excitatory amino acid transporter 2; HDAC, histone deacetylase.