1
Fig. S1
HeLa cells
ROS dye added after trypsinization
ROS dye added before trypsinization
Per
cent
age
of D
CF
flu
ores
cenc
e po
sitiv
e ce
lls (
H2O
2 pro
duct
ion)
Per
cent
age
of e
thid
ium
flu
ores
cenc
e po
sitiv
e ce
lls (
O2.- p
rodu
ctio
n)
Control
No AA
Control
No GP
Control
No AA
Control
No GP
100 101 102 103 104
FL3-H
M13.74%
45.60%
100 101 102 103 104
FL3-H
M13.54%
41.22%
100 101 102 103 104
FL3-H
M13.74%
43.34%
100 101 102 103 104
FL3-H
M13.54%
47.21%
100 101 102 103 104
FL1-H
M1
100 101 102 103 104
FL1-H
M14.35%
2.01%
5.15%
3.00%
100 101 102 103 104
FL1-H
M14.35%
33.84%
100 101 102 103 104
FL1-H
34.73%
5.15%
2
Fig. S2
a
b
U87 Cells
Ethidium, control
Ethidium, GP starvation
DCF, control
DCF, GP starvation
U87 Cells
Ethidium, control
Ethidium, AA starvation
DCF, control
DCF, AA starvation
3
No GP - + - - + - - + - - + -
No AA - - + - - + - - + - - +
NH4Cl + + + +
Time 1 h 6 h 24 h 48 h
U87 cells
Fig. S3
LC3-I
LC3-II
β-Actin
0
20
40
60
80
% C
ells
with
AV
Os
or
GF
P-L
C3
do
ts
No 3-MA
Plus 3-MA
4
Fig. S4
No GP - + - - + -
No AA - - + - - +
Autophagy AVOs GFP-LC3
Time (h) 72 h 48 h
HeLa cells
*
*
* *
* p < 0.002a
5
b
(ii)
0 1000FL3-H
M1
0 1000FL3-H
M1
ControlNo GPWort
Wort+No GP
Name M1 (%)
5.1645.135.05
10.80
Key
Control
No AA
Wort+No AA
Name M1 (%)
5.16
36.52
6.01
Key
HeLa wt HeLa wt
Fig. S4
HeLa wt
***
**
* p<0.04
HeLa SOD2
#
##
#
# p>0.05(iii)
(i)
6
0
20
40
60%
Ce
lls w
ith A
VO
s o
rG
FP
-LC
3 d
ots
Control siRNA
beclin-1 siRNA
atg-7 siRNA
**
**
** * *
* p<0.01 compared to
“control siRNA”
No GP - + - - + -
No AA - - + - - +
Autophagy AVOs GFP-LC3
Time (h) 72 h 48 h
HeLa cells
Fig. S4
c
7
HeLa cells wt SOD2 wt SOD2
Lysate Mitochondrial Cytosolic
Actin
COX2
SOD2
Fig. S5
8
Fig. S6
R - + + - - - - + + - - -
2-ME - - + + + - - - + + + -
T - - - - + + - - - - + +
Fluorescence Ethidium DCF
Time 6 h
HeLa cells
* *
**
*
*
* p<0.05
9
Fig. S7
R - + + - - - - + + - - -
2-ME - - + + + - - - + + + -
T - - - - + + - - - - + +
Fluorescence Ethidium DCF
Time 6 h
HEK293cells
a
b
R - + + - - - - + + - - -
2-ME - - + + + - - - + + + -
T - - - - + + - - - - + +
Fluorescence Ethidium DCF
Time 24 h
HEK293cells
* *
**
* p<0.05
* *
*** p<0.05
0
10
20
30
40
50
% C
ells
with
GF
P-L
C3
do
ts
10
Fig. S7
R - + + - - -
2-ME - - + + + -
T - - - - + +
Time 24 h
HeLa cells
**
**
* p<0.01c
d
R - + - - + - - + - - + -
T - - + - - + - - + - - +
2-ME - - - + + + - - - + + +
NH4Cl - +
HEK293 cells
LC3-I
LC3-IIβ-actin 0
.1
0.0
07
2 0.8
4.1
14
.2
10
.4
28
.5
45
38
.57
1.2
63
.8
11
Fig. S8
b
1 mM H2O2 - +
NH4Cl +
Time 24 h
HEK293 cells
a
LC3-II
β-Actin
LC3-I
Ethidium fluorescence DCF fluorescence
0200
Co
un
ts
100 104FL3-H
0200
Co
un
ts
100 104FL1-H
Control
1 mM H2O2 Treatment
Control
HEK293 cells
