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. Author manuscript; available in PMC: 2015 Dec 16.
Published in final edited form as: Cell Rep. 2015 Nov 19;13(9):1747–1756. doi: 10.1016/j.celrep.2015.10.064

Figure 4. Regulation of Narp by BDNF involves activation of CREB.

Figure 4

(A) Blocking transcription with actinomycin D (100nM) blocks Narp expression.

(B) Top: 300bp fragment upstream of the Narp transcription start site. Promoter fragment (yellow), Egr1 binding site (pink), CREB binding site (red), transcription start site (blue). Bottom: Outline of the luciferase reporter experiments.

(C) Luciferase induction after 3 and 6hrs of BDNF treatment. Increase in luciferase intensity with CREB-Egr1 and ΔEgr1 but not with ΔCREB or ΔCREB-Egr1. n=3, *p<0.05 ***p<0.001, mean ± SEM.

(D) 6-fold enrichment of pCREB on the Narp promoter after BDNF treatment, n=4, p<0.05, mean ± SEM. (E) EMSA confirmation of transcription factor binding. Gel shift observed in both untreated and BDNF treated nuclear extracts with labeled wild-type CREB oligo (lanes 2 and 5) but not with mutant oligo (lane 3 and 6) or excess unlabeled wild-type CREB oligo (lane 4 and 7). (F) Protein-DNA supershift in the presence of pCREB.