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. Author manuscript; available in PMC: 2016 Apr 1.
Published in final edited form as: Nat Neurosci. 2015 Aug 24;18(10):1464–1473. doi: 10.1038/nn.4095

Figure 6. Brd4 regulates synaptic receptor subunit GluA1.

Figure 6

(a) 24-hour Jq1 treatment decreases Gria1 but not Gria2 mRNA in the absence of exogenous stimulation in cultured cortical neurons. (one sample t test, for Gria1 n = 12 biological replicates, P = 6.852E-6, t = 7.959, for Gria2 n = 6 biological replicates). (b) 24 or 48-hour Jq1 treatment decreased gluA1 protein in neurons. Representative of 3 biological replicates. (c, d) GluA1 surface staining (c) and quantification (d) in neurons transfected with GFP and treated with Jq1 or the negative enantiomer for 24 hours (unpaired two-tailed t test, for control n = 33 neurons and for Jq1 n = 26 neurons from 4 biological replicates, P = 4.242E-8, t = 6.322). (e, f) Surface GluA1 staining (e) and quantification (f) in neurons transfected with GFP and either a nontargeting siRNA pool or Brd4, Brd2, or Brd3 siRNA (unpaired two-tailed t test with Bonferroni correction, n = 33 neurons for control siRNA, for Brd2 siRNA n = 31, for Brd3 siRNA n = 28, and for Brd4 siRNA n = 28 from 5 biological replicates, for control vs Brd4 P = 0.00958, t = 2.667). (g) Brd4 staining in neurons transfected with GFP and a construct expressing Brd4 under a constitutively active promoter. (h, i) GluA1 surface staining (h) and quantification (i) 2 days after transfection with GFP and Brd4 (unpaired two-tailed t test, for GFP n = 40 neurons and for GFP-Brd4 n = 35 neurons from 5 biological replicates, P = 0.00167, t = 3.264). ###, p<0.001 with univariate analysis. **, p<0.01, ***, p<0.001. ROI, region of interest. a.u. arbitrary units. h, hours. Full-length blots are presented in Supplementary Figure 11. Error bars represent standard error. Scale bar is 10 μM.