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. 2012 May 22;7(5):e37999. doi: 10.1371/journal.pone.0037999

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Lesscher et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

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A 14-3-3ζ specific shRNA sequences (1222 and 1854 bp) were cloned into pLentiLox3.7 vectors and B western blot analysis revealed that both constructs effectively reduced 14-3-3ζ protein levels in Neuro2A cells, the 1854 construct being most effective in vitro. C In situ hybridization confirmed effective knockdown of 14-3-3ζ in the CeA after infection with the 1854 shRNA expressing lentivirus. The top panels show 14-3-3ζ expression in the CeA after infection with the control or the 1854 shRNA expressing lentivirus. The bottom panels show GFP mRNA and therefore the infection site in adjacent sections. 14-3-3ζ mRNA is completely absent in the area that is infected with the 1854 shRNA expressing lentivirus.