Fig. 1.

Design and validation of the TPRT protocol. (A) Overview of TPRT, compared with ribosome profiling. (B) Changes in translation of RPs, other TOP- and TOP-like mRNAs, and non-RP internal controls, under Torin-1 treatment (100 nM, 1 h) in MDA-MB-468 cells, as measured by TPRT and ribosome profiling. Translation changes are relative to normalization factor of non-RP internal controls, and DMSO vehicle control, as described in SI Appendix, Materials and Methods. For TPRT, fold changes are derived as means over four biological replicates, each with four technical replicates; error bars denote SEs. For ribosomal profiling, fold changes derived as means of 2 biological replicates. (Inset) Direct comparison between TPRT and ribosome profiling mean fold changes in translation.