Figure 5. H4K8Ac and HDAC3 at the Fos and Nr4a2 promoters in the C57BL/6 mouse NAc are affected by conditioning with cocaine.

A) A schematic of the experimental design illustrates how C57BL/6 mice were treated with 5 mg/kg cocaine or saline and sacrificed exactly as the mice utilized for Figures 3 and 4 (fully described in Materials and Methods). B) Chromatin immunoprecipitation (ChIP) with anti-H4K8Ac antibodies isolated significantly more Fos promoter from the NAc of cocaine conditioned mice compared to saline conditioned mice. ** indicates p < 0.001. C) ChIP with anti-HDAC3 antibodies isolated significantly less Fos promoter from the NAc of cocaine conditioned mice compared to saline conditioned mice. * indicates p < 0.05. D) Anti-H4K8Ac ChIP of the Nr4a2 promoter isolated more of the promoter from the NAc of mice conditioned with cocaine compared to saline. ** indicates p < 0.001. E) Anti-HDAC3 ChIP isolated less Nr4a2 promoter from the NAc after conditioning with cocaine compared to saline. * indicates p < 0.05. For all ChIP studies, saline n = 6 mice, cocaine n = 6 mice. F) Hdac3 mRNA levels in the NAc remain unchanged after cocaine conditioning. Saline n = 5 mice, cocaine n= 5 mice, p = 0.4927. G) HDAC3 immunoreactivity from NAc proteins subjected to Western blot analyses also remains unchanged after conditioning with cocaine compared to saline. A representative image of 2 mice/group is shown within the graph. Saline n = 8 mice, cocaine n= 8 mice, p = 0.308.