Visualization of Endogenous and Exogenous Hydrogen Peroxide Using A
Lysosome-Targetable Fluorescent Probe
Dabin Kim,1 Gyoungmi Kim,1 Sang-Jip Nam,1 Jun Yin,1,2* and Juyoung Yoon1*
Contents
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UV/vis absorption spectra and fluorescence spectra of probe 1 ……………………………………………………2
Cell imaging of probe 1 ……………………………………………………………………………………………………………………………3
Mass spectra of reaction…………………………………………………………………………………………………………………………4
HPLC data……………………………………………………………………………………………………………………………………………………5
Mass and NMR Data…………………………………………………………………………………………………………………………………8
Department of Chemistry and Nano Science, Ewha Womans University, Seoul 120-750, Korea,
2
Key Laboratory of
Pesticide and Chemical Biology, Ministry of Education, College of Chemistry, Central China Normal University, Wuhan 430079,
P. R. China. *E-mail: yinj@mail.ccnu.edu.cn; jyoon@ewha.ac.kr
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1. UV/vis absorption spectra and fluorescence spectra of probe 1.
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Absorbance
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Wavelength (nm)
Figure S1. The absorption spectra of probe 1 (2 μM) with ROS (200 μM) in PBS (pH 7.4) solution
containing 1% DMF for 2 h at 25℃
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L
Absorbance
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Wavelength (nm)
Figure S2. The time-dependent absorption spectra of probe 1 (2 μM) with H2O2 (200 μM) in PBS
(pH 7.4) solution containing 1% DMF for 2 h at 25℃
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Figure S3: (a) The time-dependent fluorescence spectra of probe 1 (2 M) with H2O2 (200 M) in PBS
(pH 7.4) solution containing 1% DMF after incubation for 0-1.5 h at 25℃. Excitation wavelength = 405 nm,
excitation and emission slit widths = 3 x 5 mm.
2.
Cell imaging of probe 1
Figure S4. Probe 1 was localized to lysosomes in the HeLa cell. Confocal microscope images: (A) Probe
1 (red, ex405/em490-590 nm); (B) Bright field (gray); (C) Overlay of (A) and (B) (purple). (The cells were
incubated with 5 M probe 1 for 30 min and washed with DPBS and incubated with H2O2, ClO-, ONOOfor 30 min. Scale bar: 10 m)
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Figure S5: Probe 1 was localized to lysosomes in NIH -3T3 cell. Confocal microscope images of
probe 1: (A) Probe 1 (red, ex405/em490-590nm); (B) LysoTracker (blue, ex405/em430-455nm); (C)
Bright field (gray); (D) Overlay of (A) and (B) (purple). Left: probe 1; Middle: probe 1 + 100 μM
H2O2; Right: probe 1 + 200 μM H2O2. (The cells were incubated with 5 μM probe 1, 100 nM
Lysotracker for 30 min and washed with DPBS and incubated with H2O2 for 30 min. Scale bar: 10
μm.)
3.
Mass spectra of reaction.
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Figure S6: The FAB-MS spectra of Probe 1.
Figure S7: The FAB-MS spectrum after reaction of Probe 1 and H2O2.
4.
HPLC data.
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Figure S8: The HPLC spectra before and after reaction of Probe 1 (0.5mg/mL) and H2O2 (1 eq.).
Figure S9: The time-dependent HPLC spectra of Probe 1 (0.5mg/mL) and H2O2 (1 eq.).
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Figure S10: The time-dependent HPLC spectra of Probe 1 (0.5mg/mL) and ClO- (1 eq.).
Figure S11: The time-dependent HPLC spectra of Probe 1 (0.5mg/mL) and ONOO- (1 eq.).
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5. Mass and NMR Data
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