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. 2010 Nov 24;286(8):6641–6649. doi: 10.1074/jbc.M110.168872

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© 2011 by The American Society for Biochemistry and Molecular Biology, Inc.

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FIGURE 6. — Effect of NMD suppression on down-regulation of GPx1 mRNA after MeHg exposure. A, NMD suppression by synthetic siRNA-mediated knockdown of SMG-1 or SMG-7. Western blots of C2C12-DMPK160 transfected with the indicated synthetic siRNAs were analyzed with the indicated antibody probes. NS, non-silencing; pUpf1, anti-phospho-Upf1 antibody. B, synthetic siRNA-mediated knockdown of SMG-1 or SMG-7 results in the rescue of GPx1 mRNA down-regulation after MeHg exposure. C2C12-DMPK160 transfected with the indicated siRNAs was exposed to 0.4 μm MeHg, and total RNA was isolated 5 h or 9 h after exposure. Levels of GPx1 or β-actin mRNA were determined by quantitative real-time PCR. The histogram depicts the GPx1 mRNA normalized to the β-actin mRNA value represented as a fold-increase over NS siRNA- and non-MeHg-treated control. SMG-1- or SMG-7-knockdown cells revealed significantly higher values than NS-siRNA transfectants. Values shown are means ± S.E. of four separate experiments.