Nature - USA (2019-07-18)

reSeArCH Letter

a No stress HS Ars

YTHDF2

G3BP1

GAPDH

Tubulin

SG

Marker

Cytosol

Markers

NucCytSGSolNucCytSGSolNucCytSGSol

d

Cumulative Probability

1.00

0.75

0.50

0.25

0.00

-2 0 2

log2 fold change (stress granule mRNA/total mRNA)

1- 13

m^6 A peaks/gene

(number of genes)

Arsenite Heat shock Thapsigargin

Annotated m^6 A sites/gene

SG mRNA enrichment (log2 fold change)

2-3 kb (3856 genes) 3-4 kb (2762 genes) 4-5 kb (1831 genes) 5-6 kb (1144 genes)

0 123 +4

0

-1.0

1.0

4.0

c

Western blot

Antibody

-2 1- 0 132 -2 1- 0 132

m^6 A peaks/gene

(number of genes)

m^6 A peaks/gene

(number of genes)

0 (4742)

1 (2120)

2 (1000)

3 (449)

4+ (460)

0 (4742)

1 (2120)

2 (1000)

3 (449)

4+ (460)

0 (5262)

1 (2353)

2 (1135)

3 (502)

4+ (499)

2.0

3.0

-2.0

-3.0

-4.0

0 123 +4^0123 +4^0123 +4

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

Stress granule poly(A) RNA

b

m^6 A peaks/gene

(number of genes)

0 (2830)

1 (508)

2 (279)

3 (192)

4+ (386)

Isoxazole

Cumulative Probability

1.00

0.75

0.50

0.25

0.00

(^0123456)
log2 fold change (neuronal granule mRNA/supernatant mRNA)
e
Extended Data Fig. 4 | See next page for caption.