Live-cell single RNA imaging reveals bursts of translational frameshifting

Kenneth Lyon; Luis U. Aguilera; Tatsuya Morisaki; Brian Munsky; Timothy J. Stasevich

To confirm active translation elongation, puromycin (Sigma Aldrich) was used to release nascent chains from elongating ribosomes, leading to a rapid loss of nascent chain signal at translation sites. Bead-loaded cells with visible translation sites were imaged at a rate of one volume every seven seconds. After acquiring 16 pre-images, cells were treated with a final concentration of 100 µg/ml puromycin and continuously imaged for an additional 100 time points. As a control, the same imaging conditions were performed except that the cells were treated with vehicle (H2O). In this case, nascent chain signals All rights reserved. No reuse allowed without permission.