University of Toronto T-Space



Supplementary Data

Detecting and Imaging of SO2 Derivatives in Living Cells with Zero Cross-Talk Colorimetric Mitochondria-targeted Fluorescent Probe

Dan WuA, Shiqi RongA, Yi LiuA, Fei ZhengA, Yankun ZhaoA,Ruiwu YangB, Xiaogang DuB, Fengyan MengB, Ping ZouA and Guangtu WangA,*

A College of science, Sichuan Agricultural University, Ya’an 625014 (P. R. China).

B College of life science, Sichuan Agricultural University, Ya’an 625014 (P. R. China).

* Corresponding authors-

Email: gtwang@sicau.

Synthesis route

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Fig. S1 Synthetic route of the CA-ID-MC

Experimental

Synthetic procedures

Synthesis of the compound 4. As presented in Fig. S1, in briefly, compound 1 (1.1 mmol) was added in a solution of 2, 3, 3-trimethyl-3H-indole (1 mmol) in toluene (5 mL). The suspension was stirred at 110oC overnight under a nitrogen atmosphere, and then cooled to room temperature, and the resulting solid was collected by filtration. Then the solid was washed with cold acetone and dried under reduced pressure. The crude product was used without further purification because the purity is enough for the next synthesis.

Synthesis of the CA-ID-MC. To a solution of compound 4 (0.25 mmol) and in anhydrous ethanol, compound 3 (0.25 mmol) was added. The reaction mixture was refluxed overnight followed by evaporation of the solvent in a vacuum. Column chromatography was then used to purify the residue to yield the final product (CA-ID-MC). 1H NMR (600 MHz, CDCl3) δ 9.19 (s, 1H), 8.57 – 8.47 (m, 2H), 8.43 (d, J = 15.5 Hz, 1H), 8.33 (d, J = 15.6 Hz, 1H), 8.23 (s, 1H), 7.83 (d, J = 9.7 Hz, 1H), 7.63 – 7.51 (m, 6H), 7.46 (d, J = 7.7 Hz, 2H), 7.38 (t, J = 7.4 Hz, 1H), 7.19 (d, J = 8.6 Hz, 1H), 6.57 (s, 2H), 6.34 (t, J = 13.4 Hz, 1H), 4.40 (q, J = 7.0 Hz, 2H), 1.93 (s, 6H), 1.49 (t, J = 7.2 Hz, 3H). 13C NMR (101 MHz, DMSO) δ 176.75, 155.60, 153.52, 148.88, 138.73, 137.76, 135.92, 126.32, 125.39, 124.96, 123.98, 123.02, 122.44, 120.96, 119.90, 118.50, 117.81, 117.70, 116.59, 114.53, 112.73, 112.10, 109.56, 105.17, 104.61, 46.98, 44.47, 33.42, 23.02, 9.16. MS (ESI) m/z for C36H31BrN2O2, 523.2380, found: 523.2393 (M+).

Comparison of probes

Table S1 the comparison of fluorescent probes for SO2 derivatives.

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The performance of CA-BI-MC dissolved in methanol or DMSO

As shown in Fig. S2, a dual emission with peaks at 470 nm and 615 nm was produced when CA-BI-MC dissolved in 50% DMSO which is similar to dissolved in 50% methanol. So no matter dissolved in 50% methanol or 50% DMSO, CA-BI-MC performed similarly.

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Fig. S2 the fluorescence spectra of CA-BI-MC when dissolved in 50% methanol and 50% DMSO respectively, excited at 365 nm.

Mechanism study

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Fig. S3 (A) the 1H NMR spectrum of the mechanism study in CD3OD. (B) HRMS spectra of the mechanism study.

The time response of CA-BI-MC and CA-ID-MC

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Fig. S4 the fluorescence spectra of time response of probes towards SO32-, red line: the intensity of CA-BI-MC at 617 nm excited at 365 nm, dark line: the intensity of CA-ID-MC at 602 nm excited at 358 nm.

Data of cell toxicity

Cell viability by standard MTT assays, the experiments were repeated five times and the data are shown as mean (±S.D).

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Fig. S5 plots of the MTT assays. (A) the plot of ca-id-mc; (b) the plot of ca-bi-mc.

The experiment data of CA-ID-MC

Quantum yield: the quantum yield of CA-ID-MC was calculated to be 0.189.

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Fig. S6 (A) UV-Visible absorption spectrum of the CA-ID-MC (40 μM) upon addition of Na2SO3 solution (0-25 eq) in solution (methanol/water = 1/1, v/v), inset: the titration curve plotted with the absorbance ratio of the CA-ID-MC (A278/A520) as a function of SO32-. (B) Fluorescence titration spectra of the CA-ID-MC (40 μM) upon addition of Na2SO3 solution (0-25 eq) in solution (methanol/water = 1/1, v/v) excited at 358 nm, inset: the titration curve plotted with the intensity ratio of CA-ID-MC (I457/I602) as a function of SO32- concentration.

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Fig. S7 (A) Plot of the fluorescent intensity ratio (I457/I602) of the CA-ID-MC (40 μM) in the absent and present of SO32- (0.1 mM) to various anions and molecules (5 mM GSH and other anions, 1 mM Cys, 0.1 mM S2-) in solution (methanol/water = 1/1, v/v). (B) Plot of emission ratios (I457/I602) of CA-ID-MC (40 μM) with the increasing value of pH in water excited at 358 nm in the absent and present of SO32- (0.1 mM).

[pic]Fig. S8 HeLa cells were stained with the CA-ID-MC (50 μM) and Mito-Tracker Green (2 μM) for 30 min simultaneously. (A) Mito-Tracker Green; (B) the CA-ID-MC; (C) merged images of (A) and (B); (D) the bright field of the CA-ID-MC, insert: co-localization image across HeLa cells. Excited at 528 nm for red channel, and excited at 488 nm for Mito-Tracker Green. Bars: 10 μm.

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Fig. S9 Fluorescence images of the HeLa cells, which were cells were incubated with the CA-ID-MC (50 μM) at 37oC for 30 min in DMEM media supplemented with 10% FBS: (A) the red channel of CA-ID-MC only; (B) the green channel of the CA-ID-MC only; (C) the bright field of the CA-ID-MC only; (D) fluorescence imaging of the HeLa cells incubated with the CA-ID-MC and further incubated with SO32- (1 mM) from the red channel; (E) the green channel of (D); (F) the bright field of (D); (G) fluorescence imaging of the HeLa cells incubated with the CA-ID-MC and further incubated with SO2 donor (1 mM) and Cys (10 mM) for another 30 min from the red channel; (H) the green channel of (G); (i) the bright field of (G); R(1), R(2) and R(3) the merge of red and green channel; (J) the fluorescent intensity ratio (Fred/Fgreen) in cells. Bars: 100 μm.

Spectra

1H NMR of CA-ID-MC in CDCl3

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13C NMR of CA-ID-MC in DMSO

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1H NMR CA-BI-MC in CDCl3

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13C NMR of CA-BI-MC in CDCl3.

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HRMS of CA-ID-MC

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HRMS of CA-BI-MC

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References

[1] Huo F.; Wu Q.; Yin C.; et al. Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy, 2019, 214: 429-435.

[2] Liu A. K.; Ji R. X.; Shen S. L.; et al. New Journal of Chemistry, 2017, 41: 10096-10100.

[3] Wang L.; Li W. X.; Zhi W. J.; et al. Dyes and Pigments, 2017, 147: 357-363.

[4] Xie X. X.; Yin C. X.; Yue Y. K.; et al. Sensors and Actuators B, 2018, 147: 357-363.

[5] Xu J.; Zheng D. J.; Su M. M.; et al. Organic and Biomolecular Chemistry, 2018, 16: 8318-8324.

[6] Li Y. R.; Shi L. J.; Zhang Y. Y.; et al. Dyes and Pigments, 2018, 160: 794-798.

[7] Wang Y. F.; Meng Q. T.; Zhang R.; et al. Journal of Luminescence, 2017, 192: 297-302.

[8] Sun C.; Cao W. F.; Zhang W.; et al. Dyes and Pigments, 2019, 171: 107709.

[9] Cai F. Y.; Hou B.; Zhang S. P.; et al. Journal of Materials Chemistry B, 2019, 7: 2493-2498.

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