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. Author manuscript; available in PMC: 2018 Feb 1.
Published in final edited form as: Mol Cancer Res. 2016 Nov 29;15(2):213–224. doi: 10.1158/1541-7786.MCR-16-0247

Figure 2.

Figure 2

MYC promotes translation and total protein synthesis by augmenting mRNA cap methylation of Wnt/β-catenin signaling pathway transcripts. (A) Polysome profiling of ribosome-bound mRNA from P493-6 cells at three time points of MYC induction (0, 1, and 24 h). Data shown are from one representative sample. (B) m7G-mRNA levels of Wnt signaling transcripts in each collected fraction (1 through 6) normalized to total mRNA input and fraction 1. (0 h – top panel, 1 h – middle panel, 24 h – bottom panel) (n=3, biological). (C) Quantification of total protein synthesis detected with HRP-streptavidin (n=3, biological). (D) Western blot detecting phosphorylation status and total LRP6 and β-catenin in P493-6 cells at 0 h and 24 h. (E) Relative Wnt signaling activity measured in IMEC and IMEC:MYC cells using a TCF/LEF luciferase reporter assay (n=3, biological). Expressed as ratio (TOP Flash/FOP Flash). (F) Relative Wnt signaling activity measured in IMEC and IMEC:MYC cells following RNMT depletion with siRNA using a TCF/LEF luciferase reporter assay (n=3, biological). Expressed as ratio (TOP Flash/FOP Flash). Error bars indicate SEM and asterisks indicate significance (*p < 0.05).