Supporting Information. An Aptazyme-Gold Nanoparticle Sensor for Amplified. Molecular Probing in Living Cells

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1 Supporting Information n ptazyme-gold Nanoparticle Sensor for mplified Molecular Probing in Living Cells Yanjing Yang, Jin Huang*, Xiaohai Yang, Ke Quan, He Wang, Le Ying, Nuli Xie, Min Ou, Kemin Wang* State Key Laboratory of Chemo/iosensing and Chemometrics, College of Chemistry and Chemical Engineering, Key Laboratory for io-nanotechnology and Molecular Engineering of Hunan Province, Hunan University, Changsha 8, P. R. China. * kmwang@hnu.edu.cn;jinhuang@hnu.edu.cn Table of contents: Figure S. Characterization of unps and aptazyme-unps s Figure S. Evaluation of mounts of substrate strands on each unp S Figure S. Optimization of the ratio of substrate strands to aptazyme strands S Figure S. Kinetic study of different ratio of substrates to aptazyme s Figure S. Optimization of the concentrations of Mg S Figure S. Demonstration of the signal amplification effect of aptazyme-unp s Figure S7. Growth inhibition assay (MTT) S Figure S8. Studies of nuclease stability of the aptazyme-unp S Figure S9. ICP-MS for cellular uptake amount of the aptazyme-unps S Figure S. TEM images of cellular uptake of aptazyme-unps S Figure S. Fluorescence imaging for SMMC-77 cells S7 S

2 Supporting Figures..8 unps aptazyme-unps. bs... 7 Figure S. Characterization of unps and aptazyme-unps. () TEM micrograph of unps; () bsorption spectra of unps (black line) and aptazyme-unps (red line). The maximum optical absorption was shifted from 9 nm to nm after modification Concentration(nM) Figure S. Evaluation of mounts of substrate strands on each unp. Standard linear calibration curve of fluorescence signal against the concentration of TMR labeled substrate strands. The excitation wavelength was nm and the emission wavelength was from to nm. The error bars represent the standard deviations. S

3 F/F : : : : Figure S. Optimization of the ratio of substrate strands to aptazyme strands. F/F of the aptazyme-unps using different ratio of substrate strands to aptazyme strands after mm TP induced cleavage reaction for overnight. Error bars indicated the standard deviations of three experiments. 7 background mm TP 7 background mm TP F/F F/F Time(s) Time(s) Figure S. Kinetic study of different ratio of substrates to aptazymes. Time-dependent fluorescence response over background fluorescence with varying the ratio of substrate to aptazyme () :, () : functionalized onto the unp. s shown, the group () has higher signal turnover capability but lower reaction velocity in comparison with the group (). S

4 background mm TP Mg + [mm] Figure S. Optimization of the concentrations of Mg +. lack and gray bar denote fluorescence background and signal, respectively. Error bars indicated the standard deviations of three experiments. The result showed that the optimal concentration of Mg + was found to be mm. * - M * - M * - M 7* - M * - M * - M * - M * - M * -7 M * - M * - M * - M * - M * - M * - M * - M * - M * - M C F/F : : TP[M] Figure S. Demonstration of the signal amplification effect of aptazyme-unp. Fluorescence spectra of the aptazyme-unps sensors in the presence of various concentrations TP, with varying the ratio of substrate to aptazyme () :, () : functionalized onto the unp; (C). Comparison of the curve and detection limit between () and (). The detection limits were calculated to be µm (:) and nm (:) respectively, on the basis of the σ/slope. Error bars indicated the standard deviations of three experiments. S

5 aptazyme-unps(nm) aptazyme-unps(nm) aptazyme-unps(nm) Cell Viability (%) 8 h h 8h h Time(h) Figure S7. Growth inhibition assay (MTT). HeLa cells were incubated with with different concentrations ( nm, nm and nm) of the aptazyme-unps for h, h, 8 h and h. lack bar stands for no aptazyme-unp; light grey bar stands for the aptazyme-unp (nm); dark grey bar stands for higher concentration of the aptazyme-unp (nm). Error bars represent variations between three measurements. without DNase I with DNase I Time(s) Figure S8. Studies of nuclease stability of the aptazyme-unp. Fluorescence curves of the aptazyme-unps ( nm) in buffer with ( ) or without ( ) DNase I, as a function of time. The results showed that the introduction of DNase I did not lead to the obvious increase of fluorescence signal of the sensors. S

6 8 NO. of nanocarrier( )/cell 7 Hela SMMS-77 7 Time(h) Figure S9. ICP-MS for cellular uptake amount of the aptazyme-unps. The error bars are the standard deviations by testing samples. nucleus cytoplasm Figure S. TEM images of cellular uptake of aptazyme-unps. HeLa cells incubated with the aptazyme-unps at 7 C for h. Then the internalization and localization of the aptazyme-unps in HeLa cells was characterized by TEM. Scale bar = μm. S

7 Merge TMR Figure S. Fluorescence imaging for SMMC-77 cells. Fluorescence images and the merge of fluorescence and bright-field images of SMMC-77 cells after incubation with nm aptazyme-unp () or nm mismatched aptazyme-unp () for h at 7 C. Scale bar: μm. S7