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Results: 7

1.
Figure 1.

Figure 1. From: Monitoring eukaryotic and bacterial UDG repair activity with DNA-multifluorophore sensors.

(a) Structures of natural and unnatural nucleosides incorporated in probes; (b) structures of probes 1–3. Probes 1 and 2 are fluorogenic designs, whereas probe 3 yields a ratiometric signal.

Toshikazu Ono, et al. Nucleic Acids Res. 2013 July;41(12):e127-e127.
2.
Figure 7.

Figure 7. From: Monitoring eukaryotic and bacterial UDG repair activity with DNA-multifluorophore sensors.

Analysis by flow cytometry of signals from cells transfected with probes 4 and 7, measured 6 h post-transfection. (A) Representative histograms of pyrene emission intensity of cells transfected with no probe, probe 4 and control probe 7. (B) Averaged fold changes in fluorescence in cells transfected with probes 4 and 7 relative to cells without a probe. Error bars show standard deviations (n=3).

Toshikazu Ono, et al. Nucleic Acids Res. 2013 July;41(12):e127-e127.
3.
Figure 2.

Figure 2. From: Monitoring eukaryotic and bacterial UDG repair activity with DNA-multifluorophore sensors.

In vitro enzymatic responses of probes 1 (A), 2 (B), and 3 (C) with E. coli UDG. Time courses of fluorescence response are shown at the indicated wavelengths; insets show full spectral changes over the same time course. Conditions: 400 nM probe, UDG 1 U/ml (0.4 nM), 37°C, excitation 340 nm.

Toshikazu Ono, et al. Nucleic Acids Res. 2013 July;41(12):e127-e127.
4.
Figure 4.

Figure 4. From: Monitoring eukaryotic and bacterial UDG repair activity with DNA-multifluorophore sensors.

In vitro responses of probes 2, 4 and control 6 to human UNG2 enzyme. (A) Time courses of response of probes at 480 nm. (B) Spectral changes in response of probe 2 over the same time course. Excitation = 340 nm. Condition: [hUNG2] = 9.7 mg/ml (90 nM); [probe] = 4 μM; 37°C, excitation 340 nm.

Toshikazu Ono, et al. Nucleic Acids Res. 2013 July;41(12):e127-e127.
5.
Figure 5.

Figure 5. From: Monitoring eukaryotic and bacterial UDG repair activity with DNA-multifluorophore sensors.

Fluorescence detection and imaging of UDG activity associated with bacterial cells using probe 2. A UDG knockout strain of E. coli [BW(310)DE] was used as control; it was transfected with the AfuUDG gene, and fluorescence was observed for bacterial suspensions in solution (A) and by epifluorescence microscopy (excitation 340–380 nm, emission >400 nm) (B). Enhanced signals are seen with induction of gene expression by IPTG and at longer incubation times with the probe.

Toshikazu Ono, et al. Nucleic Acids Res. 2013 July;41(12):e127-e127.
6.
Figure 3.

Figure 3. From: Monitoring eukaryotic and bacterial UDG repair activity with DNA-multifluorophore sensors.

In vitro responses of probes 2 and 4 to hSMUG1, along with thymine-containing control probe 6. (A) Time courses of response at 480 nm. (B) Spectral changes in response of probe 2. Conditions: [hSMUG1] = 500 U/ml (400 nM), [probe] = 4 μM, 37°C, excitation 340 nm. (C) Photograph showing visible change (illumination by 365 nm UV lamp). For photograph, [hSMUG1] = 500 U/ml (400 nM), [probe 2] = 4 μM.

Toshikazu Ono, et al. Nucleic Acids Res. 2013 July;41(12):e127-e127.
7.
Figure 6.

Figure 6. From: Monitoring eukaryotic and bacterial UDG repair activity with DNA-multifluorophore sensors.

Imaging UDG activity in A253 cells by confocal fluorescence microscopy. (B and C) Images of cells treated with probes 2 and 4 (sequences shown); (A) results with thymine-containing control probe 6, and (D) positive control (fully fluorescent probe). Bright field images (E–H) of the same cells are shown below each corresponding fluorescence image. Cells were treated with 2 μM probe and with Lipofectamine as a transfection reagent for 2 h before imaging. Two-photon excitation at 710 nm.

Toshikazu Ono, et al. Nucleic Acids Res. 2013 July;41(12):e127-e127.

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