Nature - USA (2020-09-24)

Article

-2 -1 012

769-P

-3 -2 -1 01

shNC

sh1

sh2

sh4

shAGPAT3:

769-P

-2 -1 012

0.0

0.5

1.0

1.5

Relative viability

786-O

-3 -2 -1 01

786-O

-2 -1 0

sgNC

AGPAT3-sg 1

AGPAT3-sg 2

769-P

-3 -2 -1

Log 10 (M RSL3)

786-O

-2 -1 0

0.0

0.4

0.8

1.2

Relativ

ev

iability

OVCAR-8

ij

-1 01

769-P

0.0

0.4

0.8

1.2

Relativ

ev

iabilit

y

Log 10 (M ML210)

sgNC

AGPAT3-sg1

AGPAT3-sg2

f g

Log 10 (M ML210) Log 10 (M RSL3)

-Log

10

p adj.

-2 -1 012

0.0

0.5

1.0

1.5

2.0

2.5

Log 2 (AGPAT 3 -sg / sgNC)

769-P

otherphospholipids

PUFA-ePE/ePC

diacyl-PUFA-PE/PC

l

-2 -1 0

Log 10 (M RSL3)

OVCAR-8-AGPAT3-sg1

+Vector

+mAgpat3-WT

+mAgpat3-E176A

k

CEG C A G

175 176 177

WT mAgpat3 cDNA

AGPAT3-sg2

sgNC

OVCAR-8 10 52 .5 1.25 0.625 0.313 0.156 0.078 0.039 Vehicle

(day 5)

ML210(M)

175 176 177

mAgpat3-E176A cDNA

e

h

Log 10 (M RSL3) Log 10 (M RSL3)

a

50

37

β-ACTIN

75

ACSL4

12 NC

AGPS-sg

12 NC

PEX12-sg

12 NC

PEX3-sg

12 NC

FAR1-sg

12

PEX10-sg

75

50

LPCAT3

786-O

MW(KDa)

β-ACTIN

12 NC

AGPS-sg

12 NC

AGPAT3-sg

12 NC

PEX3-sg

12

FAR1-sg

OVCAR-8

75

ACSL4

75

50

LPCAT3

50

37

MW(KDa)

b

c

ACSL 4

FAR1

AGPS

GAPDH

Dox:

OVCAR-8

FAR1-sg2

AGPS-sg2:

:

PEX3-sg1:

PEX1 0 -sg1:

sgNC:

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-ACSL4-sg1TetOn

-1 01

0.0

0.4

0.8

1.2

-1 01 -1 01 -1 01

sgNC, -dox

sgNC, -dox +Fer-1

sgNC,ACSL 4 -sg1TetOn, +dox

sg-GOI,ACSL 4 -sg1TetOn, -dox

sg-GOI,ACSL 4 -sg1TetOn, +dox

sg-GOI,ACSL 4 -sg1TetOn, -dox +Fer-1

Log 10 (M ML210)

Relative viability

AGPS-sg 2 FAR1-sg (^2) PEX3-sg1 PEX1 0 -sg 1
d
-1 01
0.0
0.5
1.0
1.5
Relative viabilit
y
Log 10 (M ML210)
shNCsh1sh2sh
0.0 (^3) sh4
0.5
1.0
1.5
AGPAT3
(786-O)
Relative
mRNA
leve
l
p=0.043p=0.047p=0.09p=0.006
shNCsh1sh2sh3sh4
AGPAT3
(769-P)
p=0.142
p=0.01
1
p=0.0048p=0.0014
Extended Data Fig. 7 | See next page for caption.