Nature - USA (2019-07-18)

Letter reSeArCH

a b c

G3BP1 G3BP1

Merge Merge

Arsenite (1 h) Heat Shock (30 min)

d

DF1

Merge

DF3

Merge

Heat Shock (30 min)Heat Shock (30 min)

TIAR TIAR

DF1

Merge

Arsenite (1 h)

TIAR

DF3

Merge

Arsenite (1 h)

TIAR

f

TIAR

DF2

Merge

e

Arsenite (1 h)

TIAR

DF2

Merge

g

Heat shock (45 min) Thapsigargin (1.5 h)

TIAR

DF2

Merge

TIAR TIAR

DF2 DF2

Merge Merge

Total cells

j

Stress condition

Heat shock, 10 min

Heat shock, 45 minNIH3T30 30

Heat shock, 30 min mES 038

Arsenite, 30 min mES 043

Arsenite, 60 minHEK293 029

Arsenite, 60 minU2OS0 25

Arsenite, 60 minNIH3T30 35

Arsenite, 60 min mES 045

Thapsigargin, 1.5hrNIH3T30 35

Thapsigargin, 3hr mES 026

Cell type DF2 in nucleus

mES 042

DMSO

Heat Shock (30 min)

Puromycin Actinomycin D

DF2

Arsenite (1 h) Arsenite (1 h)

i

No stress Arsenite (1 h)

DF2-NeonGreen DF2-NeonGreen

h

DF2-

DF2

GAPDH

CRISPR/Cas9 CRISPR/Cas9

anti-YTHDF2 anti-NeonGreen

65kDa

100kDa

37kDa

l

No stress Heat shock, 30 min Arsenite, 1 h

m^6 A m^6 A m^6 A

A

C

U U U

A A

C C

G G G

mES cells mES cells

HEK293 U2OS NIH3T3

n.s. (p = 0.51)

NeonGreen

k

Control

HeatShoc

k

Arsenite

0.00

0.02

0.04

0.06

m

6 A/(A+C+U)

n.s. (p = 0.70)

mES cells

Extended Data Fig. 2 | See next page for caption.