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nature research | reporting summary
October 2018#2861354, 1:100), rabbit anti-G3BP1 (13057-2-AP, Proteintech, Lot #00047654, 1:100), mouse anti-ATXN2 (Clone 22) (611378,
BD Biosciences, Lot #7341666, 1:100), rabbit anti-YTHDF1 (17479-AP, Proteintech, Lot #00044547, 1:100), rabbit anti-YTHDF2
(24744-1-AP, Proteintech, Lot #00053880, 1:100), rabbit anti-YTHDF3 (ab103328, Abcam, Lot #GR35115-39, 1:100), rabbit anti-
IgG Alexa Fluor 594 (A11012, Invitrogen, Lot #1933366, 1:1000), mouse anti-IgG Alexa Fluor 488 (A11001, Invitrogen, Lot
#1939600, 1:1000).The following antibodies were used for immunoblotting experiments: rabbit anti-YTHDF2 (ARP67917_P050, Aviva Systems
Biology, Lot #QC38405-43182, 1:1000), mouse anti-GAPDH (SC-365062, Santa Cruz Biotechnology, Lot #A2816, 1:5000), rabbit
anti-IgG HRP (NA934V, GE Healthcare, Lot #16677077, 1:10000), mouse anti-IgG HRP (NA931V, GE Healthcare, Lot #16814909,
1:10000).Validation Antibodies used for IF:
rabbit anti-TIAR (5137S, Cell Signaling Technology) - manufacturer has validated use for IF in mouse and human
mouse anti-TIAR (Clone 6) (610352, BD Biosciences) - manufacturer has validated use for IF in mouse and human
mouse anti-Edc4 (H-12) (sc-376382, Santa Cruz Biotechnology) - manufacturer has validated use for IF in mouse and human
mouse anti-Puromycin clone 12D10 (MABE343, Millipore Sigma) - manufacturer has validated use for IF in mouse and human
rabbit anti-G3BP1 (13057-2-AP, Proteintech) - manufacturer has validated use for IF in mouse and human
mouse anti-ATXN2 (Clone 22) (611378, BD Biosciences) - manufacturer has validated use for IF in human. ATXN2 is a well-
characterized stress granule marker, and this antibody has been used in IF studies previously in monkey (Nonis et al., 2008) and
rat (Nonhoff et al., 2007). We found ATXN2 co-localized well with G3BP1 (a known stress granule marker) in both human cells
and mouse embryonic stem cells (Extended Data 2a). ATXN2 localization to SGs is similar to what has been observed in previous
stress granule studies (Kaehler et al. 2012) in human cells.
rabbit anti-YTHDF1 (17479-AP, Proteintech) - manufacturer has validated use for IF in mouse and human
rabbit anti-YTHDF2 (24744-1-AP, Proteintech) - manufacturer has validated use for IF in mouse and human
rabbit anti-YTHDF3 (ab103328, Abcam) - manufacturer has validated use for IHC in human. A previous publication validated the
use of this antibody for IF in mouse (Yu et al., 2017 - Dynamic m6A modification regulates local translation of mRNA in axons)Antibodies used for WB:
rabbit anti-YTHDF2 (ARP67917_P050, Aviva Systems Biology) - manufacturer has validated use for WB in human
mouse anti-GAPDH (SC-365062, Santa Cruz Biotechnology) - manufacturer has validated use for WB in humanEukaryotic cell lines
Policy information about cell lines
Cell line source(s) Mettl14 knockout mouse embryonic stem cells were a gift from J. Hanna and S. Geula (Weizmann Institute of Science).
HEK293 (ATCC CRL-11268), U2OS (ATCC HTB-96), and NIH3T3 (ATCC CRL- 1658) were obtained from ATCC.Authentication Mettl14 knockout mES cells were previously described in Geula, et. al. 2015. HEK293, U2OS, and NIH3T3 cells were not
authenticated.Mycoplasma contamination Mycoplasma staining was routinely tested with Hoechst staining. No mycoplasma were detected at any point in any cell line
used in this study.Commonly misidentified lines
(See ICLAC register)No commonly misidentified cell lines were used in this study.