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. Author manuscript; available in PMC: 2017 May 5.
Published in final edited form as: Mol Cell. 2016 Apr 21;62(3):335–345. doi: 10.1016/j.molcel.2016.03.021

Figure 2. METTL3 enhances translation.

Figure 2

(A–B) METTL3 knockdown in HeLa cells (A) Western blot using indicated antibodies. (B) q.RT-PCR of indicated mRNAs normalized to β-Actin mRNA. Relative ratio (fold-change) obtained in the presence of shGFP was set to 1. Data are presented as ±SEM. n=3. (C) Western blot analysis of METTL3 in nuclear and cytoplasmic fractions using β-Tubulin (cytoplasmic, Cy) and Fibrillarin (nuclear, Nc) as controls. (D–E) Cytoplasmic extracts from control or METTL3-depleted cells were subjected to sucrose grandient centrifugation. (D) Polysome-fractionated samples analyzed by Western blot using the indicated antibodies and RT-PCR performed with α-[32P]-dCTP. (E) Relative levels of EGFR or TAZ mRNAs in each ribosome fraction were quantified and normalized to RCN2 mRNA and plotted as a percentage of the total. Data are from three independent polysome-profiling experiments. Error bars = mean ±SEM, n=3.