1. The initial targeting of Sec61b mRNA to the ER is partially dependent on ribosomes and translation.
The initial targeting of Sec61b mRNA to the ER is partially dependent on ribosomes and translation. U2OS cells were pretreated with control medium (Ctrl) or HHT for 15 min, then microinjected with plasmids containing GFP–Sec61b and allowed to express mRNAs for 2 h in the presence of medium with or without HHT. The cells were then extracted with digitonin, fixed and stained with FISH probe against the GFP-coding sequence, and imaged. (A) Representative images, with each row representing a single field of view imaged for GFP–Sec61b mRNA (mRNA) and GFP fluorescence (GFP). (B) Quantification of the fluorescence intensities of mRNAs in the ER and nucleus of extracted cells. Each bar represents the mean±s.e.m. of three independent experiments, each experiment consisting of at least 30 cells. Scale bar: 20 µm.
Xianying A. Cui et al. J Cell Sci 2015;128:
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