0
200
400
600
800
1000
1200
1400
1600
Control
hsp-6 hsp-60 dnj-10 hsp-3 hsp-4
hsp-16.2 C12C8.1
hsp-1 F44E5.4
GFP Fluorescence (RFU)
**
**
0
10
20
30
40
50
60
70
80
90
100
hsp-4 spliced
xbp-1
hsp-70
(C12C8.1)
hsp-16.2 hsp-17 hsp-1
mRNA Fold induction after hsp-6 RNAi ER UPR Cytosolic HSR
A
B
hsp-16.2p::GFP;CF512
Control(EV)hsp-6 hsp-60 dnj-10 hsp-3 hsp-4 hsp-16.2 C12C8.1 hsp-1 F44E5.4
hsp-16.2p::GFP
Mitochondrial RNAi ER RNAi Cytosolic RNAi
Control(EV) EV:hsp-6 hsp-6:clpp-1hsp-6:dve-1hsp-6:hsf-1
Double RNAi with hsp-6
C
hsp-16.2p::GFP
200
400
600
800
1000
1200
Control
EV:
hsp-6
clpp-1:hsp-6
dve-1:hsp-6 hsf-1:hsp-6
GFP Fluorescence (RFU)^0
**
hsp-16.2p::GFP;CF512
Figure 1. Knockdown of Mitochondrial HSP70,hsp-6, Induces Cytosolic Heat Shock Response via UPRmt
(A)hsp-6andhsp-1RNAi-induced cytosolic small heat shock protein expression.hsp-16.2p::GFP induction was measured by COPAS biosorter (bottom)
(mean±SD of three biological repeats; p%0.01). EV, empty vector control RNAi.
(B) qPCR of UPRERand cytosolic HSR genes afterhsp-6RNAi (mean±SD of three biological repeats).
(C) Cytosolic HSR induced byhsp-6RNAi was dependent on bothhsf-1and UPRmtcomponents.hsp-16.2p::GFP induction was measured by COPAS biosorter
(bottom) (mean±SD of three biological repeats; p%0.001).
See alsoFigure S1and Table S1.
Cell 166 , 1539–1552, September 8, 2016 1541