REAGENT or RESOURCE	SOURCE	IDENTIFIER
Chemicals, Peptides, and Recombinant Proteins
DMEM	GIBCO	Cat# 31966021
Leibovitz’s L15 medium	GIBCO	Cat# 21083-027
Penicillin-Streptomycin	GIBCO	Cat# 15140-122
Fetal Bovine Serum (FBS)	Sigma-Aldrich	Cat# F7524
Doxycycline	Sigma-Aldrich	Cat# D9891-1G
Opti-MEM	Sigma-Aldrich	Cat# 11058-021
FuGENE 6 Transfection Reagent	Promega	Cat# E231A
Lipofectamine RNAi-MAX	Invitrogen	Cat# 13778-075
Polyethylenimine	Polysciences Inc	Cat# 23966
Zeocin	Invitrogen	Cat# R25001
Trimethoprim	Sigma-Aldrich	Cat# T7883-5G
TRIsure	Bioline	Cat# Bio-38033
Bioscript Reverse Transcriptase	Bioline	Cat# Bio-27036
Halo-TMR ligand	Promega	Cat# G8252
RNase Inhibitor	New England Biolabs (NEB)	Cat# M0307L
Polybrene	Santa Cruz Biotechnology, Inc	Cat# sc-134220
Critical Commercial Assays
iQ SYBR Green SuperMix	Bio-Rad	Cat# 1708885
Experimental Models: Cell Lines
Human U2OS cells	Tanenbaum lab	Cat# HTB-96
HEK293T cells	Tanenbaum lab	Cat# CRL-3216
Oligonucleotides
siRNA targeting sequence-UPF1:GCAGUUCCGCUCCAUUUUGAU	Dharmacon	N/A
siRNA targeting sequence-XRN1: AGAUGAACUUACCGUAGAAUU	Dharmacon	N/A
siRNA targeting sequence-SMG6: GGGUCACAGUGCUGAAGUAUU	Dharmacon	N/A
See Table S1 for all primers used for RT-qPCRs.	This study	N/A
Recombinant DNA
See Table S1 for all plasmids used in the paper	This study	N/A
Software and Algorithms
ImageJ	NIH	https://imagej.nih.gov/ij/
Graphpad Prism 7	GraphPad Software Inc	https://www.graphpad.com/scientific-software/prism/
MATLAB R2012b	The Mathworks, Inc.	https://nl.mathworks.com/products/matlab.html
Micromanager for microscope control	Micro-Manager 1.4.22	https://micro-manager.org
NIS elements 5.11.01	Nikon	https://www.microscope.healthcare.nikon.com/en_EU/products/software
TransTrack (MATLAB)	Boersma et al., 2018	https://github.com/TanenbaumLab
RiboFitter (R)	Boersma et al., 2018	https://github.com/TanenbaumLab
Other
96-well glass bottom imaging plates-(Matriplates)	Brooks Life Science Systems	Cat# MGB096-1-2-LG-L
Deposited Data
Raw data of imaging experiments	Mendeley data	https://doi.org/10.17632/bw255hcw7h.1