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. 2020 Aug 18;48(17):9550–9570. doi: 10.1093/nar/gkaa671

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© The Author(s) 2020. Published by Oxford University Press on behalf of Nucleic Acids Research.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 6. — Fos eRNA1 is necessary for Fos mRNA expression in neurons. (A) Anti-sense oligonucleotide (ASO) targeting of Fos eRNA1 for 24 h decreased both eRNA1 and Fos mRNA (unpaired t-test t(10) = 20.69, P < 0.0001 and t(10) = 5.739, P = 0.0002), but did not alter eRNA levels from other Fos enhancers (unpaired t-test for Gapdh mRNA t(10) = 0.9696, P = 0.3551; eRNA2 t(10) = 0.8608, P = 0.4095; eRNA3 t(10) = 1.014, P = 0.3346). (B) Fos mRNA targeting ASOs decreased Fos mRNA (t(10) = 5.198, P = 0.0004) with no significant effect on eRNA levels (unpaired t-test for Gapdh mRNA t(10) = 3.744, P = 0.0038; eRNA1 t(10) = 2.056, P = 0.0668; eRNA2 t(10) = 0.6508, P = 0.5298; eRNA3 t(10) = 1.679, P = 0.124). (C) Fos eRNA1 ASO pretreatment for 24 h prior to 1 h Veh treatment or KCl stimulation reduced induction of eRNA1 (top) and mRNA (bottom) when compared to a scrambled ASO control (n = 9 per group, two-way ANOVA for eRNA1 F(1,32) = 154.4 P < 0.0001, for mRNA F(1,32) = 5.267, P = 0.0284). Data expressed as mean ± s.e.m. Multiple comparisons, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.