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In vivo NO generation and anti-cancer effect of PMT system. a Schematic illustration of the NO responsive element contained sensing plasmid and its in vitro NO responsiveness ( n = 3). b In vivo NO generation of PMT system in 4T1 Nrf2-luc tumor-bearing mice. The imaging was performed 24 h after the i.v. injection of E. coli , CCN@ E. coli (10 8 CFU mL −1 in 200 μL saline), and CCN (5 mg mL −1 in 200 μL saline). c <t>MRI</t> images for detecting both in vitro and in vivo NO generation with the assistance of NO responsive contrast <t>agent</t> <t>Fe-MGD</t> ( n = 3). The in vivo imaging was performed 24 h after the i.v. injection of CCN@ E. coli (10 8 CFU mL −1 in 200 μL saline). d Imaging apoptotic response in vivo with annexin V-Cy5.5 (Red: CCN@ E. coli , Green: apoptotic cancer cells). The in vivo imaging was performed 48 h after the i.v. injection of CCN@ E. coli (10 8 CFU mL −1 in 200 μL saline). e Semiquantitative analysis of relationship among intratumoral NO concentration (evaluated by the circle area), bacteria count, and signal intensity of apoptotic cells ( n = 3). f Schematic diagram of the process of in vivo anti-cancer therapy. Intravenous injection of E. coli , CCN@ E. coli (10 8 CFU mL −1 in 200 μL saline), CCN (5 mg mL −1 in 200 μL saline) and phosphate buffer solution treated group (PBS) were performed on the 1st and 7th days ( n = 6). Light irradiation was performed 24 h after the injection ( > 630 nm, 30 mW cm −2 , 15 min). g In vivo anti-cancer effect of PMT in 4T1 tumor-bearing mice ( n = 6). h In vivo anti-cancer effect of PMT in CT26 tumor-bearing mice ( n = 6). i H&E staining of 4T1 tumor after 14 days of PMT (Scale bar: 100 μm). j TUNEL staining of 4T1 tumor after 14 days of PMT (Scale bar: 100 μm). k Three-dimensional fluorescence imaging for visualizing the co-localization of CCN@ E. coli and apoptotic region within the tumor. Significance between every two groups was calculated using unpaired two-tailed Student’s t -test. * P < 0.05, ** P < 0.01, *** P < 0.001. The mean values and S.D. are presented
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In vivo NO generation and anti-cancer effect of PMT system. a Schematic illustration of the NO responsive element contained sensing plasmid and its in vitro NO responsiveness ( n = 3). b In vivo NO generation of PMT system in 4T1 Nrf2-luc tumor-bearing mice. The imaging was performed 24 h after the i.v. injection of E. coli , CCN@ E. coli (10 8 CFU mL −1 in 200 μL saline), and CCN (5 mg mL −1 in 200 μL saline). c <t>MRI</t> images for detecting both in vitro and in vivo NO generation with the assistance of NO responsive contrast <t>agent</t> <t>Fe-MGD</t> ( n = 3). The in vivo imaging was performed 24 h after the i.v. injection of CCN@ E. coli (10 8 CFU mL −1 in 200 μL saline). d Imaging apoptotic response in vivo with annexin V-Cy5.5 (Red: CCN@ E. coli , Green: apoptotic cancer cells). The in vivo imaging was performed 48 h after the i.v. injection of CCN@ E. coli (10 8 CFU mL −1 in 200 μL saline). e Semiquantitative analysis of relationship among intratumoral NO concentration (evaluated by the circle area), bacteria count, and signal intensity of apoptotic cells ( n = 3). f Schematic diagram of the process of in vivo anti-cancer therapy. Intravenous injection of E. coli , CCN@ E. coli (10 8 CFU mL −1 in 200 μL saline), CCN (5 mg mL −1 in 200 μL saline) and phosphate buffer solution treated group (PBS) were performed on the 1st and 7th days ( n = 6). Light irradiation was performed 24 h after the injection ( > 630 nm, 30 mW cm −2 , 15 min). g In vivo anti-cancer effect of PMT in 4T1 tumor-bearing mice ( n = 6). h In vivo anti-cancer effect of PMT in CT26 tumor-bearing mice ( n = 6). i H&E staining of 4T1 tumor after 14 days of PMT (Scale bar: 100 μm). j TUNEL staining of 4T1 tumor after 14 days of PMT (Scale bar: 100 μm). k Three-dimensional fluorescence imaging for visualizing the co-localization of CCN@ E. coli and apoptotic region within the tumor. Significance between every two groups was calculated using unpaired two-tailed Student’s t -test. * P < 0.05, ** P < 0.01, *** P < 0.001. The mean values and S.D. are presented
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In vivo NO generation and anti-cancer effect of PMT system. a Schematic illustration of the NO responsive element contained sensing plasmid and its in vitro NO responsiveness ( n = 3). b In vivo NO generation of PMT system in 4T1 Nrf2-luc tumor-bearing mice. The imaging was performed 24 h after the i.v. injection of E. coli , CCN@ E. coli (10 8 CFU mL −1 in 200 μL saline), and CCN (5 mg mL −1 in 200 μL saline). c <t>MRI</t> images for detecting both in vitro and in vivo NO generation with the assistance of NO responsive contrast <t>agent</t> <t>Fe-MGD</t> ( n = 3). The in vivo imaging was performed 24 h after the i.v. injection of CCN@ E. coli (10 8 CFU mL −1 in 200 μL saline). d Imaging apoptotic response in vivo with annexin V-Cy5.5 (Red: CCN@ E. coli , Green: apoptotic cancer cells). The in vivo imaging was performed 48 h after the i.v. injection of CCN@ E. coli (10 8 CFU mL −1 in 200 μL saline). e Semiquantitative analysis of relationship among intratumoral NO concentration (evaluated by the circle area), bacteria count, and signal intensity of apoptotic cells ( n = 3). f Schematic diagram of the process of in vivo anti-cancer therapy. Intravenous injection of E. coli , CCN@ E. coli (10 8 CFU mL −1 in 200 μL saline), CCN (5 mg mL −1 in 200 μL saline) and phosphate buffer solution treated group (PBS) were performed on the 1st and 7th days ( n = 6). Light irradiation was performed 24 h after the injection ( > 630 nm, 30 mW cm −2 , 15 min). g In vivo anti-cancer effect of PMT in 4T1 tumor-bearing mice ( n = 6). h In vivo anti-cancer effect of PMT in CT26 tumor-bearing mice ( n = 6). i H&E staining of 4T1 tumor after 14 days of PMT (Scale bar: 100 μm). j TUNEL staining of 4T1 tumor after 14 days of PMT (Scale bar: 100 μm). k Three-dimensional fluorescence imaging for visualizing the co-localization of CCN@ E. coli and apoptotic region within the tumor. Significance between every two groups was calculated using unpaired two-tailed Student’s t -test. * P < 0.05, ** P < 0.01, *** P < 0.001. The mean values and S.D. are presented
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In vivo NO generation and anti-cancer effect of PMT system. a Schematic illustration of the NO responsive element contained sensing plasmid and its in vitro NO responsiveness ( n = 3). b In vivo NO generation of PMT system in 4T1 Nrf2-luc tumor-bearing mice. The imaging was performed 24 h after the i.v. injection of E. coli , CCN@ E. coli (10 8 CFU mL −1 in 200 μL saline), and CCN (5 mg mL −1 in 200 μL saline). c <t>MRI</t> images for detecting both in vitro and in vivo NO generation with the assistance of NO responsive contrast <t>agent</t> <t>Fe-MGD</t> ( n = 3). The in vivo imaging was performed 24 h after the i.v. injection of CCN@ E. coli (10 8 CFU mL −1 in 200 μL saline). d Imaging apoptotic response in vivo with annexin V-Cy5.5 (Red: CCN@ E. coli , Green: apoptotic cancer cells). The in vivo imaging was performed 48 h after the i.v. injection of CCN@ E. coli (10 8 CFU mL −1 in 200 μL saline). e Semiquantitative analysis of relationship among intratumoral NO concentration (evaluated by the circle area), bacteria count, and signal intensity of apoptotic cells ( n = 3). f Schematic diagram of the process of in vivo anti-cancer therapy. Intravenous injection of E. coli , CCN@ E. coli (10 8 CFU mL −1 in 200 μL saline), CCN (5 mg mL −1 in 200 μL saline) and phosphate buffer solution treated group (PBS) were performed on the 1st and 7th days ( n = 6). Light irradiation was performed 24 h after the injection ( > 630 nm, 30 mW cm −2 , 15 min). g In vivo anti-cancer effect of PMT in 4T1 tumor-bearing mice ( n = 6). h In vivo anti-cancer effect of PMT in CT26 tumor-bearing mice ( n = 6). i H&E staining of 4T1 tumor after 14 days of PMT (Scale bar: 100 μm). j TUNEL staining of 4T1 tumor after 14 days of PMT (Scale bar: 100 μm). k Three-dimensional fluorescence imaging for visualizing the co-localization of CCN@ E. coli and apoptotic region within the tumor. Significance between every two groups was calculated using unpaired two-tailed Student’s t -test. * P < 0.05, ** P < 0.01, *** P < 0.001. The mean values and S.D. are presented
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In vivo NO generation and anti-cancer effect of PMT system. a Schematic illustration of the NO responsive element contained sensing plasmid and its in vitro NO responsiveness ( n = 3). b In vivo NO generation of PMT system in 4T1 Nrf2-luc tumor-bearing mice. The imaging was performed 24 h after the i.v. injection of E. coli , CCN@ E. coli (10 8 CFU mL −1 in 200 μL saline), and CCN (5 mg mL −1 in 200 μL saline). c <t>MRI</t> images for detecting both in vitro and in vivo NO generation with the assistance of NO responsive contrast <t>agent</t> <t>Fe-MGD</t> ( n = 3). The in vivo imaging was performed 24 h after the i.v. injection of CCN@ E. coli (10 8 CFU mL −1 in 200 μL saline). d Imaging apoptotic response in vivo with annexin V-Cy5.5 (Red: CCN@ E. coli , Green: apoptotic cancer cells). The in vivo imaging was performed 48 h after the i.v. injection of CCN@ E. coli (10 8 CFU mL −1 in 200 μL saline). e Semiquantitative analysis of relationship among intratumoral NO concentration (evaluated by the circle area), bacteria count, and signal intensity of apoptotic cells ( n = 3). f Schematic diagram of the process of in vivo anti-cancer therapy. Intravenous injection of E. coli , CCN@ E. coli (10 8 CFU mL −1 in 200 μL saline), CCN (5 mg mL −1 in 200 μL saline) and phosphate buffer solution treated group (PBS) were performed on the 1st and 7th days ( n = 6). Light irradiation was performed 24 h after the injection ( > 630 nm, 30 mW cm −2 , 15 min). g In vivo anti-cancer effect of PMT in 4T1 tumor-bearing mice ( n = 6). h In vivo anti-cancer effect of PMT in CT26 tumor-bearing mice ( n = 6). i H&E staining of 4T1 tumor after 14 days of PMT (Scale bar: 100 μm). j TUNEL staining of 4T1 tumor after 14 days of PMT (Scale bar: 100 μm). k Three-dimensional fluorescence imaging for visualizing the co-localization of CCN@ E. coli and apoptotic region within the tumor. Significance between every two groups was calculated using unpaired two-tailed Student’s t -test. * P < 0.05, ** P < 0.01, *** P < 0.001. The mean values and S.D. are presented
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In vivo NO generation and anti-cancer effect of PMT system. a Schematic illustration of the NO responsive element contained sensing plasmid and its in vitro NO responsiveness ( n = 3). b In vivo NO generation of PMT system in 4T1 Nrf2-luc tumor-bearing mice. The imaging was performed 24 h after the i.v. injection of E. coli , CCN@ E. coli (10 8 CFU mL −1 in 200 μL saline), and CCN (5 mg mL −1 in 200 μL saline). c <t>MRI</t> images for detecting both in vitro and in vivo NO generation with the assistance of NO responsive contrast <t>agent</t> <t>Fe-MGD</t> ( n = 3). The in vivo imaging was performed 24 h after the i.v. injection of CCN@ E. coli (10 8 CFU mL −1 in 200 μL saline). d Imaging apoptotic response in vivo with annexin V-Cy5.5 (Red: CCN@ E. coli , Green: apoptotic cancer cells). The in vivo imaging was performed 48 h after the i.v. injection of CCN@ E. coli (10 8 CFU mL −1 in 200 μL saline). e Semiquantitative analysis of relationship among intratumoral NO concentration (evaluated by the circle area), bacteria count, and signal intensity of apoptotic cells ( n = 3). f Schematic diagram of the process of in vivo anti-cancer therapy. Intravenous injection of E. coli , CCN@ E. coli (10 8 CFU mL −1 in 200 μL saline), CCN (5 mg mL −1 in 200 μL saline) and phosphate buffer solution treated group (PBS) were performed on the 1st and 7th days ( n = 6). Light irradiation was performed 24 h after the injection ( > 630 nm, 30 mW cm −2 , 15 min). g In vivo anti-cancer effect of PMT in 4T1 tumor-bearing mice ( n = 6). h In vivo anti-cancer effect of PMT in CT26 tumor-bearing mice ( n = 6). i H&E staining of 4T1 tumor after 14 days of PMT (Scale bar: 100 μm). j TUNEL staining of 4T1 tumor after 14 days of PMT (Scale bar: 100 μm). k Three-dimensional fluorescence imaging for visualizing the co-localization of CCN@ E. coli and apoptotic region within the tumor. Significance between every two groups was calculated using unpaired two-tailed Student’s t -test. * P < 0.05, ** P < 0.01, *** P < 0.001. The mean values and S.D. are presented
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In vivo NO generation and anti-cancer effect of PMT system. a Schematic illustration of the NO responsive element contained sensing plasmid and its in vitro NO responsiveness ( n = 3). b In vivo NO generation of PMT system in 4T1 Nrf2-luc tumor-bearing mice. The imaging was performed 24 h after the i.v. injection of E. coli , CCN@ E. coli (10 8 CFU mL −1 in 200 μL saline), and CCN (5 mg mL −1 in 200 μL saline). c MRI images for detecting both in vitro and in vivo NO generation with the assistance of NO responsive contrast agent Fe-MGD ( n = 3). The in vivo imaging was performed 24 h after the i.v. injection of CCN@ E. coli (10 8 CFU mL −1 in 200 μL saline). d Imaging apoptotic response in vivo with annexin V-Cy5.5 (Red: CCN@ E. coli , Green: apoptotic cancer cells). The in vivo imaging was performed 48 h after the i.v. injection of CCN@ E. coli (10 8 CFU mL −1 in 200 μL saline). e Semiquantitative analysis of relationship among intratumoral NO concentration (evaluated by the circle area), bacteria count, and signal intensity of apoptotic cells ( n = 3). f Schematic diagram of the process of in vivo anti-cancer therapy. Intravenous injection of E. coli , CCN@ E. coli (10 8 CFU mL −1 in 200 μL saline), CCN (5 mg mL −1 in 200 μL saline) and phosphate buffer solution treated group (PBS) were performed on the 1st and 7th days ( n = 6). Light irradiation was performed 24 h after the injection ( > 630 nm, 30 mW cm −2 , 15 min). g In vivo anti-cancer effect of PMT in 4T1 tumor-bearing mice ( n = 6). h In vivo anti-cancer effect of PMT in CT26 tumor-bearing mice ( n = 6). i H&E staining of 4T1 tumor after 14 days of PMT (Scale bar: 100 μm). j TUNEL staining of 4T1 tumor after 14 days of PMT (Scale bar: 100 μm). k Three-dimensional fluorescence imaging for visualizing the co-localization of CCN@ E. coli and apoptotic region within the tumor. Significance between every two groups was calculated using unpaired two-tailed Student’s t -test. * P < 0.05, ** P < 0.01, *** P < 0.001. The mean values and S.D. are presented

Journal: Nature Communications

Article Title: Optically-controlled bacterial metabolite for cancer therapy

doi: 10.1038/s41467-018-03233-9

Figure Lengend Snippet: In vivo NO generation and anti-cancer effect of PMT system. a Schematic illustration of the NO responsive element contained sensing plasmid and its in vitro NO responsiveness ( n = 3). b In vivo NO generation of PMT system in 4T1 Nrf2-luc tumor-bearing mice. The imaging was performed 24 h after the i.v. injection of E. coli , CCN@ E. coli (10 8 CFU mL −1 in 200 μL saline), and CCN (5 mg mL −1 in 200 μL saline). c MRI images for detecting both in vitro and in vivo NO generation with the assistance of NO responsive contrast agent Fe-MGD ( n = 3). The in vivo imaging was performed 24 h after the i.v. injection of CCN@ E. coli (10 8 CFU mL −1 in 200 μL saline). d Imaging apoptotic response in vivo with annexin V-Cy5.5 (Red: CCN@ E. coli , Green: apoptotic cancer cells). The in vivo imaging was performed 48 h after the i.v. injection of CCN@ E. coli (10 8 CFU mL −1 in 200 μL saline). e Semiquantitative analysis of relationship among intratumoral NO concentration (evaluated by the circle area), bacteria count, and signal intensity of apoptotic cells ( n = 3). f Schematic diagram of the process of in vivo anti-cancer therapy. Intravenous injection of E. coli , CCN@ E. coli (10 8 CFU mL −1 in 200 μL saline), CCN (5 mg mL −1 in 200 μL saline) and phosphate buffer solution treated group (PBS) were performed on the 1st and 7th days ( n = 6). Light irradiation was performed 24 h after the injection ( > 630 nm, 30 mW cm −2 , 15 min). g In vivo anti-cancer effect of PMT in 4T1 tumor-bearing mice ( n = 6). h In vivo anti-cancer effect of PMT in CT26 tumor-bearing mice ( n = 6). i H&E staining of 4T1 tumor after 14 days of PMT (Scale bar: 100 μm). j TUNEL staining of 4T1 tumor after 14 days of PMT (Scale bar: 100 μm). k Three-dimensional fluorescence imaging for visualizing the co-localization of CCN@ E. coli and apoptotic region within the tumor. Significance between every two groups was calculated using unpaired two-tailed Student’s t -test. * P < 0.05, ** P < 0.01, *** P < 0.001. The mean values and S.D. are presented

Article Snippet: The relaxivity of (MGD) 2 -Fe(II)-NO was measured on a 7.0 T MRI (BioSpec 70/20USR).

Techniques: In Vivo, Plasmid Preparation, In Vitro, Imaging, Injection, Saline, In Vivo Imaging, Concentration Assay, Bacteria, Irradiation, Staining, TUNEL Assay, Fluorescence, Two Tailed Test