CELF1 or HuR was silenced by transfection with a specific siRNA as described previously (Zou et al., 2010 (link); Xiao et al., 2011 (link)). The siRNAs specifically targeting mRNAs encoding CELF1 (siCUGBP1) or HuR (siHuR) and control-siRNA (C-siRNA) were purchased from Santa Cruz. For each 60-mm cell culture dish, 15 μl of the 20 μM stock duplex siCELF1, siHuR, or C-siRNA was used. Forty-eight hours after transfection using Lipofectamine, cells were harvested for analysis.
Protocol detail
CELF1 and HuR silencing protocol
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Corresponding Organization :
Other organizations : University of Maryland, Baltimore, Veterans Health Administration, Institute on Aging, National Institute on Aging, National Institutes of Health
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Variable analysis
independent variables
- Silenced expression of CELF1 or HuR mRNA using specific siRNAs (siCUGBP1 or siHuR, respectively)
- Measured outcomes after 48 hours of transfection
- Cell culture dish (60-mm)
- Transfection method (Lipofectamine)
- Control-siRNA (C-siRNA)
- Positive control: Cells transfected with siCUGBP1 or siHuR to silenced CELF1 or HuR expression, respectively.
- Negative control: Cells transfected with C-siRNA.
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