Cell - 8 September 2016

CD

hsp-16.2p::GFP (with hsp-6 RNAi)

Control Brain CM C16 C20 C22 C24

hsp-16.2p::GFP

Control Heart CL C14 C18

A

Control(EV) EV: hsp-6 hsp-6: crls-1

Nonyl Acridine Orange Staining

RNAi

B

Control(EV) EV: hsp-6 hsp-6: crls-1

hsp-16.2p::GFP

ppppppppppppppp p RNAi

E

Sphingomyelin T10B11.2

Ceramide-1-phosphate

Serine + Palmitoyl CoA

Sphinganine

3-Ketosphinganine

Dihydroceramide

Ceramide

Sphingosine

sptl-1,2,3

hyl-1,2 and lagr-1

asm-1,2,3

Ceramidase

Cardiolipin W02F12.2, F27E5.1, K11D2.2

Cardiolipin

F

RNAi hsp-16.2p::GFP

induction

hsp-16.2p ::GFP suppression

(Double RNAi with hsp- 6

RNAi)

sptl-1 Yes n/a

sptl-2 Yes n/a

sptl-3 Yes n/a

hyl-1 Yes n/a

hyl-2 Yes n/a

lagr-1 No n/a

asm-2 Yes n/a

asm-3 Yes n/a

ZK856.18 (asm-2 ortholog) Yes n/a

T10B11.2 No Yes

W02F12.2 No Yes

F27E5.1 No Yes

K11D2.2 No Yes

0

5000

10000

15000

20000

25000

30000

35000

40000

EV sptl-1 sptl-2 sptl-3 hyl-1 hyl-2 lagr-1

asm-2 ZK856.18 asm-3

Peak GFP Fluorescence (RFU)

hsp-16.2p::GFP (peak)

***

*

*

* *

** ****

**

0

100

200

300

400

500

600

700

800

EV T10B11.2 K11D2.2 F27E5.1 W02F12.2

GFP Fluorescence (RFU)

hsp-16.2p::GFP (double RNAi)

***

**

** **

Double RNAi with hsp-6 RNAi

(legend on next page)

1546 Cell 166 , 1539–1552, September 8, 2016