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. 2018 Nov 22;19(12):3716. doi: 10.3390/ijms19123716

Figure 2.

Figure 2

Expression of IEGs required de novo messenger RNA (mRNA) transcription and translation. (a) Primary neurons at 15 DIV co-treated with Bic/4-AP (50µM/2.5mM) and the translation inhibitor anisomycin A (Aniso, 25 µM) or the transcription inhibitor actinomycin D (Actino, 10 µM) for 2 h were subjected to immunoblotting with anti-Arc, c-Fos, EgrI, and actin antibodies. Results are representative of at least four experiments. Results demonstrate that both translation and transcription are required for the expression of IEGs following synaptic activation (SA). (b–d) mRNA levels of Arc, c-Fos, and EgrI were determined by quantitative PCR in primary neurons at 15 DIV treated with Bic/4-AP (50µM/2.5mM, 30 min) after 20 min with or without pre-treatment with the AMPK inhibitor Compound C (Cc, 10 µM). Results show mean ± SD (n = 4–6). One-way ANOVA followed by Bonferroni’s post-hoc test were used for evaluation of statistical significance, *** p < 0.001.