Cell - 8 September 2016

A

acetyl CoA

malonyl CoA

saturated fatty acylCoAs

monounsaturated fatty acylCoAs

triglycerides

storage/export

CPT

mitochondrial

ß-oxidation

pod-2

fasn-1

fat-5, fat-6, fat-7

Perhexiline (PHX)

B

0

200

400

600

800

1000

1200

1400

Control EV:hsp-6 hsp-6:pod-2hsp-6:fasn-1

GFP Fluorescence (RFU)

**

* **

0

100

200

300

400

500

600

700

Control dve-1 hsf-1 clpp-1 atfs-1

GFP Fluorescence (RFU)

Control

PHX

**

C

EV: hsp-6 hsp-6:pod-2 hsp-6:fasn-1

hsp-16.2p::GFP

Double RNAi with hsp-6

Control (EV) dve-1 hsf-1 clpp-1 atfs-1 RNAi

hsp-16.2p::GFP

DMSO (Control)

PHX treatment

Control (EV) dve-1 hsf-1 clpp-1 atfs-1 RNAi

hsp-16.2p::GFP

hsp-16.2p::GFP

Figure 4. Reducing Fat Synthesis Blocks Cytosolic Response, and Inhibiting CPT Activity Induces Cytosolic Stress Response
(A) Diagram showingC. elegansgenes involved in the fat storage pathway.pod-2, acyl-CoA carboxylase;fasn-1, fatty acid synthase;fat-5,fat-6, andfat-7,
delta-9 fatty acid desaturase; CPT, carnitine palmitoyltransferase.
(B) Knocking down two enzymes involved in fat synthesis inhibited cytosolic response. (Continued fromFigure 1C; note that the image of control worms are from
Figure 1C.)hsp-16.2p::GFP reporter induction was measured by COPAS biosorter (bottom) (mean±SD of three biological repeats; *p%0.05, p%0.01).
(C) CPT inhibitor PHX-treated hsp-16.2p::GFP;CF512 reporter worms showed elevated levels of GFP that were inhibited byhsf-1and UPRmtcomponents. GFP
induction was measured by COPAS biosorter (right) (mean±SD of three biological repeats; p%0.01).
See alsoFigure S4.

Cell 166 , 1539–1552, September 8, 2016 1545