Extended Data Fig. 2: Validation experiments after knockdown of decay machinery.
From: Short poly(A) tails are protected from deadenylation by the LARP1–PABP complex
a. Schematic of Tet-On inducible TOP TRICK reporter. Reporter mRNAs are only transcribed in the presence of doxycycline. SV40 poly(A) signal on its 3′ UTR was targeted for measuring poly(A)-tail length by Hire-PAT assay. b. Global poly(A) length distributions measured by mTAIL-seq upon depletion of decay factors. c. Poly(A) length distributions of pulse-expressed reporter mRNAs measured by Hire-PAT assay upon depletion of decay factors at various time points post-wash. d. Steady-state poly(A) length distribution of endogenous mRNAs measured by Hire-PAT assay upon depletion of decay factors.
