Fig. 7: The presence of one 8-oxoG adduct in the ORF of an mRNA leads to accelerated rate of decay in HEK293 cells.
From: Oxidation and alkylation stresses activate ribosome-quality control
a Schematic showing the steps used to make mRNA reporters with 8-oxoG in the ORF. b Ethidium-bromide-stained PAGE gel used to follow the construction of the mRNA showing that they migrate as would be predicted based on their size. c Microscope images of HEK293 cells electroporated with capped and uncapped eGFP mRNA used to assess electroporation efficiency. Shown are the brightfield, and GFP- and DAPI-fluorescence signals. Scale bar is 50 μm. d Northern analysis to measure the decay of the indicated reporters and the control GFP mRNAs post electroporation. Reporters were visualized directly by exposing the nylon transfer to a phosphorimager screen. GFP and GAPDH were visualized using a radiolabeled DNA probe. At the bottom is the ethidium-bromide stained agarose gel. The half-lives were determined from duplicates (±SD).
