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GAS5 interacts with eIF4E.

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posted on 08.09.2014, 03:11 by Guangzhen Hu, Zhenkun Lou, Mamta Gupta

(A) GAS5 mRNA was detected by RT-PCR after RNA-IP using eIF4E and IgG antibodies. The RNA-IP was repeated for two times with similar results. (B) The polysome and non-polysome fractions, as shown by the profile of the absorbance at 254 nm, were separated by sucrose gradient centrifugation in HEK-293T cells. The experiments were repeated three times. (C) The abundance of GAS5 lncRNA in the polysome and non-polysome fractions was measured by Q-PCR and normalized with total GAS5 mRNA in all the fractions. Bars represent mean ±SD from 3 replicates. Experiment was repeated three times, and a representative experiment is shown. (D) Protein levels of eIF4E and eIF4G in the polysome and non-polysome fractions were detected by western blot.

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