OPEN ACCESS
sensors
ISSN 1424-8220
www.mdpi.com/journal/sensors
Supplementary Information
A mRNA-Responsive G-Quadruplex-Based Drug Release System.
Sensors 2015, 15, 9388-9403
Hidenobu Yaku 1, Takashi Murashima 2, Daisuke Miyoshi 2,* and Naoki Sugimoto 2,3
1
2
3
Advanced Research Division, Panasonic Corporation, 3-4 Hikaridai, Seika-cho, Soraku-gun,
Kyoto 619-0237, Japan; E-Mail: [email protected]
Faculty of Frontiers of Innovative Research in Science and Technology (FIRST),
Konan University, 7-1-20 Minatojima-Minamimachi, Chuo-ku, Kobe 650-0047, Japan;
E-Mails: [email protected] (T.M.); [email protected] (N.S.)
Frontier Institute for Biomolecular Engineering Research (FIBER), Konan University,
7-1-20 Minatojima-Minamimachi, Chuo-ku, Kobe 650-0047, Japan
* Author to whom correspondence should be addressed; E-Mail: [email protected];
Tel: +81-78-303-1426; Fax: +81-78-303-1495.
Abs. at 690 nm
2
0.9
0.8
0.7
0.6
0.5
0.4
0.3
0.2
0.1
0
0 100 200 300 400 500 600
HteloDNA (μM)
Figure S1. Change in absorbance at 690 nm of 100 μM Cu-APC with 0–500 μM
Htelo-DNA in a buffer containing 50 mM MES-LiOH (pH 7.0), 100 mM KCl, and 10 mM
MgCl2 at 25 °C.
A
B
0.12
1
0.1
0.8
0.6
Abs. 405
Abs. 424
0.08
HteloDNA
0.06
dsDNA
0.04
0.4
HteloDNA
dsDNA
0.2
0.02
0
0
-0.02
-0.2
0
5e-5
50
1e-4
100 1.5e-4
150
DNA
(M)
DNA / µM
2e-4
200
2.5e-4
250
00
5e-5
50
1e-4
100 1.5e-4
150
DNA
(M)
DNA / µM
2e-4
200
2.5e-4
250
Figure S2. (A) Absorbance of 1.0 µM TMPyP4 at 424 nm (A) and 12.5 µM Hemin
at 405 nm (B) with 0-250 µM of HteloDNA and 0–250 µM of dsDNA
(5'-AGAAGAGAAAGA-3'/5'-TCTTTCTCTTCT-3'). Before measurement, the sample was
heated at 80 °C for 2 min, gently cooled at 2 °C·min−1. All measurements were carried out in a
buffer containing 50 mM MES-LiOH (pH 7.0), 100 mM KCl, and 10 mM MgCl2 at 25 °C.
3
A
00wt%
wt% PEG
PEG200
200
TMPyP4 (mM)
/ µM
Cu-APC
TMPyP4
Cu-APC
TMPyP4
(mM)
1.2
1.2
0.1 0.3
0.3 11 3 10
00 0.1
1.0
1.0
TRAP products
Relativeactivity
activity
Relative
0.1 0.3
0.3 1
10
1 33 10
00 0.1
20 Bwt% PEG 200
0.80
0.8
0.60
0.6
0.40
0.4
0.20
0.2
0.0
0
I.C.→
I.C.→
I.C.→
0
2
2 10
-6
-6
-6
4
6
4 10
6 10
TMPyP4
(M)
TMPyP4 /
8
8 10
-6
-5
1 10
10
µM
Figure S3. (A) Electrophoresis result of the two-step TRAP assay with 0–10 µM of
TMPyP4. I.C. indicates the internal control for PCR amplification; (B) Relative activity of
telomerase with 0–10 µM of TMPyP4. The relative activity value of 1 corresponds to the
positive control, namely without TMPyP4.
A
0 wt%PEG 200
20Bwt% PEG 200
Hemin (mM)
/ µM
Hemin
Hemin (mM)
10 30100
30100 200
00 33 10
0 3 10 30100 200
1.2
1.2
Relative activity
TRAP products
1.0
1.0
0.80
0.8
0.60
0.6
0.40
0.4
0.20
0.2
I.C.→
I.C.→
0.0
0
00
-5
5 50
10
0.0001
100
0.00015
150
0.0002
200
Hemin / µM
Figure S4. (A) Electrophoresis result of the two-step TRAP assay with 0–200 µM of
Hemin. I.C. indicates the internal control for PCR amplification; (B) Relative activity of
telomerase with 0–200 µM of Hemin. The relative activity value of 1 corresponds to the
positive control, namely without Hemin.
4
A
B
0.35
0.35
250
0.3
0.25
Absorbance
0.25
Absorbance
250
0.3
GDDC-L10
(µM)
0.2
0.15
0.1
0.2
0.15
GDDC-L15
(µM)
0.1
0
0.05
0
0.05
0
500
550
600
650
700
750
0
500
800
550
Wavelength / nm
C
650
700
750
800
750
800
D
0.35
0.35
250
0.3
250
0.3
0.25
Absorbance
0.25
Absorbance
600
Wavelength / nm
GDDC-L20
(µM)
0.2
0.15
0.1
0.2
0.15
GDDC-L30
(µM)
0.1
0
0.05
0
0.05
0
500
550
600
650
700
750
800
Wavelength / nm
0
500
550
600
650
700
Wavelength / nm
E
1.2
Normalized Abs. 690
1
0.8
GDDC-L10
GDDC-L15
GDDC-L20
GDDC-L30
0.6
0.4
0.2
0
00
50
5e-5
100
1e-4
150
1.5e-4
200
2e-4
250
2.5e-4
DNA
/ µM
DNA
conc.
(M)
Figure S5. Visible absorbance spectra of 2.5 µM CuAPC with 0–250 µM of GDDC-L10 (A);
GDDC-L15 (B); GDDC-L20 (C); and GDDC-L30 (D). Before measurement, the samples
were heated at 80 °C for 2 min, gently cooled at 2 °C·min−1; (E) Normalized absorbance of
2.5 µM CuAPC at 690 nm with 0–250 µM of each GDDC candidate. All measurements
were carried out in a buffer containing 50 mM MES-LiOH (pH 7.0), 100 mM KCl, and
10 mM MgCl2 at 25 °C.
© 2015 by the authors; licensee MDPI, Basel, Switzerland. This article is an open access article
distributed under the terms and conditions of the Creative Commons Attribution license
(http://creativecommons.org/licenses/by/4.0/).