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. 2014 Apr 11;42(9):5765–5775. doi: 10.1093/nar/gku225

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© The Author(s) 2014. 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/3.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 4. — Knockdown of endogenous UBE3A disrupts circadian rhythms in mammalian cells. (A) Representative raw (left) and detrended (right) bioluminescence traces of Bmal1::Luc NIH3T3 cells treated with shRNA #1 (top) and #2 (bottom) against Ube3a. (B) Representative traces of Per2::luc SW1353 cells treated with human Ube3a SiRNA constructs at 10 (mid-grey) and 20 nM (black) doses. Scrambled SiRNA was used as control. (C) Left panels: circadian amplitude analysis (n = 3, shRNA #1 P < 0.01; shRNA #2 P < 0.001; RNAi 10 nM, P < 0.05; RNAi 20 nM, P < 0.01). Right panel: circadian period analysis (n = 3, t-tests: RNAi 10 nM, P < 0.05; RNAi 20 nM, P < 0.05). Note: the rapid dampening of circadian oscillation in shRNA-treated NIH3T3 cells prevented proper evaluation of periodicity. *P < 0.05; **P < 0.01; ***P < 0.001.