Nature - USA (2020-09-24)

Article

c

d

050 100 150

0

20

40

60

Relative gain in

ox-BODIPY

-C1

1 signal

050 100 150050 100 150 050 100 150

Vehicle

C18 (plasm)-C20:4 PE

C18:0 - C20:4 PE

Lip- 1

AGPS-sg2 FAR1-sg2 PEX3-sg1 AGPAT3-sg1

Time (min)

050100 150200 250

0

20

40

60

Time (min)

Relative gain in

ox-BODIPY-

C1

1 signal

Vehicle

C18:0 - C20:4 PE

C18 (plasm)-C20:4 PE

Lip-1

050 100 150 200 250

0

50

100

150

200

Time (min)

Relative gain in

ox-BODIPY

-C1

1 signal

Vehicle

C18:0 - C20:4 PC

C18 (Plasm)-C20:4 PC

Lip-1

Lipid peroxidation time course (786-O) (1h delay)Lipid peroxidation time course (786-O) (1h delay)

a

04080120 160

0

50

100

150

200

250

Relative gain in

ox-BODIPY

-C11 signal

Vehicle

C18:0-C20:4 PE

C18(plasm)-C20:4 PE

Lip-1

04080 120 160

Time (min)

Lipid peroxidation time course (786-O)

Vehicle

C18:0-C20:4 PC

C18(plasm)-C20:4 PC

Lip-1

-1.5 -1.0-0.5 0.00.5 1.01.5

0

1

2

3

4

5

Log 2 (ML210 / DMSO)

-Log

10

adj.

p

786-O lipidomics

PUFA-ePE/ePC

C36:5 ePC42:11C ePE Diacyl-PUFA-PE/PC

C38:6 ePE

C38:4 PE

C36:4 PE

C40:6 PE

C38:6 PE

C22:0 LPE

C18:0 LPE

C20:4 LPE

f

Time (min)

b Lipid peroxidation time course (786-O)

0

50

100

150

200

250

Relative gain in

ox-BODIPY

-C11 signal

e

Extended Data Fig. 9 | Polyunsaturated plasmalogens promote lipid
peroxidation in GPX4-inhibited cells. a, Quantification of time-lapse imaging
of lipid peroxidation levels reported by BODIPY-C11 oxidation in 786-O cells
co-treated with ML210 and indicated PE nanoparticles or Lip-1. n = 3
biologically independent samples. b, Quantification of time-lapse imaging of
lipid peroxidation levels reported by BODIPY-C11 oxidation in 786-O cells
co-treated with ML210 and indicated PC nanoparticles or Lip-1. n = 3
biologically independent samples. c, Quantification of time-lapse imaging of
lipid peroxidation levels reported by BODIPY-C11 oxidation in 786-O cells
expressing the indicated sgRNAs treated with ML210 and indicated PE
nanoparticles or Lip-1. n = 2 biologically independent samples. Nanoparticles
were added at the same time as ML210. d, Quantification of time-lapse imaging
of lipid peroxidation levels reported by BODIPY-C11 oxidation in 786-O cells
treated with ML210 and indicated PE nanoparticles or Lip-1. n = 3 biologically

independent samples. Nanoparticles were added 1 h after ML210

administration (indicated by the arrow, note a 10 min time is deduced for

reagent and equipment handling). e, Quantification of time-lapse imaging of

lipid peroxidation levels reported by BODIPY-C11 oxidation in 786-O cells

treated with ML210 and indicated PC nanoparticles or Lip-1. n = 3 biologically

independent samples. Nanoparticles were added 1 h after ML210

administration (indicated by the arrow, note a 10 min time is deduced for

reagent and equipment handling). f, Volcano plots showing the lipidomic

analysis of 786-O cells treated with ML210 or DMSO for 90 min. n = 3

biologically independent samples. Two tailed Student’s t-test. Multiple-testing

adjustment was performed using the Benjamini–Hochberg method. For

viability curves and lipid peroxidation time-lapse imaging quantification, data

are mean ± s.d.