Figure 3.

Single-molecule imaging of translation and mRNA decay in the cytoplasm of live cells. (A) Schematic representation of translation RNA imaging by coat protein knockoff (TRICK) for measuring the first round of translation by loss of PP7 label from the coding sequence during the first round of translation. (B) Representative image of TRICK experiment showing untranslated (PP7 and MS2; white) and translated (MS2-only; magenta) messenger RNAs (mRNAs). (C) Schematic representation of 3′-RNA end accumulation during turnover (TREAT) for measuring mRNA degradation using viral pseudoknots to stabilize a decay intermediate. (D) Representative image of TREAT experiment showing intact transcripts (PP7 and MS2; white) and stabilized 3′ end decay intermediates (MS2-only; magenta) arising from Xrn1-mediated degradation.