0
200
400
600
800
1000
1200
1400
Control (EV) pod-2 fasn-1 haf-1
GFP Fluorescence (RFU)
Control
Heat Shock
A
D
hsp-6p::GFP hsp-60p::GFP
0
100
200
300
400
500
600
700
hsp-6p::GFP hsp-60p::GFP
GFP Fluorescence (RFU)
Control
PHX
*
**
B
0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
0.9
DMSO PHX
Triglyceride (mM)
C **
0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
0.9
1
Control (EV) EV:hsp-6 hsp-6:pod-2 hsp-6:fasn-1
Triglyceride (mM)
*
0
20
40
60
80
100
120
140
160
180
EV+DMSO hsp-6 +DMSO EV+PHX hsp-6 +PHX
OCR (pmol\min\10worms)
*
* *
E
DMSO PHX DMSO PHX
hsp-16.2p::GFP
(^0) 0 20406080100
100
200
300
400
mthsp70 siRNA
(^0020406080100)
100
200
300
400
Control siRNA
Time (minutes)
OCR (pmoles/min)
BSA
PALM
Oligo FCCP Rtn/AA
Oligo FCCP Rtn/AA
BSA
PA L M
siRNA: Ctrl mthsp70
mtHSP70
Tubulin
F
Figure S4. Reducing Fat Synthesis Blocks the Cytosolic Response, and Inhibiting CPT Activity Induces the Cytosolic Stress Response,
Related toFigure 4
(A) Triglyceride content was measured after indicated RNAi treatment (paired t test, mean±SD of three biological repeats, *p%0.05).
(B) Knocking down fat synthesis genespod-2andfasn-1had a specific inhibitory effect on mitochondrial to cytosolic signaling while they had no effect on heat
shock response.hsp-16.2p::GFP reporter induction was measured by COPAS biosorter after RNAi and heat shock (mean±SD of three biological repeats).
(C) Triglyceride content was measured after PHX treatment (paired t test, mean±SD of three biological repeats, **p%0.01).
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