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. Author manuscript; available in PMC: 2018 Jun 1.
Published in final edited form as: Cell. 2017 Jun 1;169(6):1051–1065.e18. doi: 10.1016/j.cell.2017.05.022

Figure 7.

Figure 7

PKM2 is enriched at ER ribosomes and localizes mRNAs to the ER.

A, Quantitative-MS experiment to characterize PKM2 containing ribosomes.

Scatter plot with normalized log2 heavy/light ratios comparing eL36 and PKM2-enriched ribosomes (n=2). Mean 0.61; S.D. 0.50; cut off values for enriched proteins was 2.5 S.D. from the mean and is shown as the grey line lines. See Table S6. Green denotes ER-related components.

B, Subcellular ER-ribosome enrichment. eL31 is tagged endogenously with Avitag at the C terminus. ER-or cytoplasmic- biotin ligase is expressed from an inducible promoter. ER-biotin ligase is attached to the Sec61 Beta protein and cytoplasmic biotin ligase contains a nuclear export signal (NES). PKM2 enrichment is shown relative to known ER resident proteins.

C, Subcellular localization of PKM2 iCLIP targets. The fraction of mRNAs within subcellular fractions normalized to total mRNA are shown. Each fraction value is initially determined by normalizing to an exogenous spike-in RNA control. Data are mean and S.D. of two biological replicates. CYT, cytosol; ER, Endoplasmic reticulum; NUC, nucleus; CSK, cytoskeleton.