f nucleatum Atcc Search Results


  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 95
    ATCC f nucleatum atcc 10953
    Effect of serum on viability of A. oris (Ao), V. parvula (Vp), F. <t>nucleatum</t> <t>ATCC</t> 10953 (Fn) and P. gingivalis 381 (Pg). Planktonic growth of microorganisms was evaluated in their appropriate nutrient-rich medium supplemented with 10% native human serum
    F Nucleatum Atcc 10953, supplied by ATCC, used in various techniques. Bioz Stars score: 95/100, based on 245 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/f nucleatum atcc 10953/product/ATCC
    Average 95 stars, based on 245 article reviews
    Price from $9.99 to $1999.99
    f nucleatum atcc 10953 - by Bioz Stars, 2020-08
    95/100 stars
      Buy from Supplier

    97
    ATCC f nucleatum atcc 23726
    PMSF inhibits the proteolytic activity of F. <t>nucleatum</t> . A, F. nucleatum FDC 364. B, F. nucleatum ATCC 25586. C, F. nucleatum 12230. D, F. nucleatum ATCC 23726. M, Molecular weight markers. Presented data are of representative zymograms.
    F Nucleatum Atcc 23726, supplied by ATCC, used in various techniques. Bioz Stars score: 97/100, based on 387 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/f nucleatum atcc 23726/product/ATCC
    Average 97 stars, based on 387 article reviews
    Price from $9.99 to $1999.99
    f nucleatum atcc 23726 - by Bioz Stars, 2020-08
    97/100 stars
      Buy from Supplier

    99
    ATCC f nucleatum atcc 25586
    (a and b) RANKL expression in PDL cells stimulated by F. <t>nucleatum</t> <t>ATCC</t> 25586 and/or CTSH at 1 day (a) and 3 days (b). (c and d) OPG expression in PDL cells stimulated by F. nucleatum ATCC 25586 and/or CTSH at 1 day (c) and 3 days (d). (e and f) RANKL/OPG mRNA ratio in PDL cells stimulated by F. nucleatum ATCC 25586 and/or CTSH at 1 day (e) and 3 days (f). *Significant difference compared to other groups ( P
    F Nucleatum Atcc 25586, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 687 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/f nucleatum atcc 25586/product/ATCC
    Average 99 stars, based on 687 article reviews
    Price from $9.99 to $1999.99
    f nucleatum atcc 25586 - by Bioz Stars, 2020-08
    99/100 stars
      Buy from Supplier

    84
    ATCC f nucleatum atcc 23726 nana
    (a and b) RANKL expression in PDL cells stimulated by F. <t>nucleatum</t> <t>ATCC</t> 25586 and/or CTSH at 1 day (a) and 3 days (b). (c and d) OPG expression in PDL cells stimulated by F. nucleatum ATCC 25586 and/or CTSH at 1 day (c) and 3 days (d). (e and f) RANKL/OPG mRNA ratio in PDL cells stimulated by F. nucleatum ATCC 25586 and/or CTSH at 1 day (e) and 3 days (f). *Significant difference compared to other groups ( P
    F Nucleatum Atcc 23726 Nana, supplied by ATCC, used in various techniques. Bioz Stars score: 84/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/f nucleatum atcc 23726 nana/product/ATCC
    Average 84 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    f nucleatum atcc 23726 nana - by Bioz Stars, 2020-08
    84/100 stars
      Buy from Supplier

    84
    ATCC × 107 f nucleatum atcc 23726
    (a and b) RANKL expression in PDL cells stimulated by F. <t>nucleatum</t> <t>ATCC</t> 25586 and/or CTSH at 1 day (a) and 3 days (b). (c and d) OPG expression in PDL cells stimulated by F. nucleatum ATCC 25586 and/or CTSH at 1 day (c) and 3 days (d). (e and f) RANKL/OPG mRNA ratio in PDL cells stimulated by F. nucleatum ATCC 25586 and/or CTSH at 1 day (e) and 3 days (f). *Significant difference compared to other groups ( P
    × 107 F Nucleatum Atcc 23726, supplied by ATCC, used in various techniques. Bioz Stars score: 84/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/× 107 f nucleatum atcc 23726/product/ATCC
    Average 84 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    × 107 f nucleatum atcc 23726 - by Bioz Stars, 2020-08
    84/100 stars
      Buy from Supplier

    88
    ATCC wild type f nucleatum atcc 23726
    F. nucleatum <t>ATCC</t> 23726 accelerates tumor growth and progression of lung metastasis. a Experimental scheme: AT3 breast cancer cell line was injected to the mammary fat pad of 6–7-week-old C57BL/6 mice. When tumor size reached 500 mm 3 , PBS vehicle (V), 5 × 10 7 F. nucleatum ATCC 23726 only (726), 5 × 10 7 F. nucleatum ATCC 23726 with metronidazole (726+MTZ.), metronidazole only (MTZ.), or 5 × 10 7 Fap2-deficient F. nucleatum K50 were injected into the tail vein. On days 2–3 post inoculation, mice were injected twice a day with metronidazole (M) or with PBS vehicle (V). On days 4–7 post inoculation, mice were injected once a day with metronidazole or with PBS. Eight days after inoculation, mice were sacrificed, and tumor and lungs harvested. Breast cancer tissues were weighed, and lung metastases were counted. b Tumor weights normalized as the percentage compared with the average tumor weight in the PBS group (vehicle control) in each experiment (set as 100% tumor weight). Each symbol represents one mouse. Presented results are of three independent experiments. **** p = 3.48 × 10 −5 , ** p = 0.002. Holm’s correction, one-tailed Mann–Whitney. Vehicle n = 21, MTZ. n = 17, K50 n = 7, 726 n = 19, 726+MTZ. n = 12. c Representative tumors post-harvest. d Representative image of lung metastases. Scale bar, 500 µm in the large image, 250 µm in the indicated inset. e Lung metastasis rank [number of metastases per lung area (pixel 2 )] calculated as percentage compared with the average metastasis rank in the vehicle control group of each experiment (set as 100% metastasis rank). Each symbol represents one mouse. Presented results are of three independent experiments. **** p = 4.08 × 10 −5 , * p = 0.0177, n.s.: non-significant, Holm’s correction, one-tailed Mann–Whitney. Vehicle n = 20, MTZ. n = 17, K50 n = 7, 726 n = 18, 726+met n = 12. f Images taken by fluorescence binocular of lung metastases in four mice implanted with GFP-expressing AT3 cells as described above. Two tumor-bearing mice were inoculated with F. nucleatum and two were treated by PBS vehicle. Scale bar, 1000 µm. Source data are provided as a Source data file.
    Wild Type F Nucleatum Atcc 23726, supplied by ATCC, used in various techniques. Bioz Stars score: 88/100, based on 13 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/wild type f nucleatum atcc 23726/product/ATCC
    Average 88 stars, based on 13 article reviews
    Price from $9.99 to $1999.99
    wild type f nucleatum atcc 23726 - by Bioz Stars, 2020-08
    88/100 stars
      Buy from Supplier

    86
    ATCC f nucleatum atcc 23727 attachment
    F. nucleatum <t>ATCC</t> 23726 accelerates tumor growth and progression of lung metastasis. a Experimental scheme: AT3 breast cancer cell line was injected to the mammary fat pad of 6–7-week-old C57BL/6 mice. When tumor size reached 500 mm 3 , PBS vehicle (V), 5 × 10 7 F. nucleatum ATCC 23726 only (726), 5 × 10 7 F. nucleatum ATCC 23726 with metronidazole (726+MTZ.), metronidazole only (MTZ.), or 5 × 10 7 Fap2-deficient F. nucleatum K50 were injected into the tail vein. On days 2–3 post inoculation, mice were injected twice a day with metronidazole (M) or with PBS vehicle (V). On days 4–7 post inoculation, mice were injected once a day with metronidazole or with PBS. Eight days after inoculation, mice were sacrificed, and tumor and lungs harvested. Breast cancer tissues were weighed, and lung metastases were counted. b Tumor weights normalized as the percentage compared with the average tumor weight in the PBS group (vehicle control) in each experiment (set as 100% tumor weight). Each symbol represents one mouse. Presented results are of three independent experiments. **** p = 3.48 × 10 −5 , ** p = 0.002. Holm’s correction, one-tailed Mann–Whitney. Vehicle n = 21, MTZ. n = 17, K50 n = 7, 726 n = 19, 726+MTZ. n = 12. c Representative tumors post-harvest. d Representative image of lung metastases. Scale bar, 500 µm in the large image, 250 µm in the indicated inset. e Lung metastasis rank [number of metastases per lung area (pixel 2 )] calculated as percentage compared with the average metastasis rank in the vehicle control group of each experiment (set as 100% metastasis rank). Each symbol represents one mouse. Presented results are of three independent experiments. **** p = 4.08 × 10 −5 , * p = 0.0177, n.s.: non-significant, Holm’s correction, one-tailed Mann–Whitney. Vehicle n = 20, MTZ. n = 17, K50 n = 7, 726 n = 18, 726+met n = 12. f Images taken by fluorescence binocular of lung metastases in four mice implanted with GFP-expressing AT3 cells as described above. Two tumor-bearing mice were inoculated with F. nucleatum and two were treated by PBS vehicle. Scale bar, 1000 µm. Source data are provided as a Source data file.
    F Nucleatum Atcc 23727 Attachment, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/f nucleatum atcc 23727 attachment/product/ATCC
    Average 86 stars, based on 5 article reviews
    Price from $9.99 to $1999.99
    f nucleatum atcc 23727 attachment - by Bioz Stars, 2020-08
    86/100 stars
      Buy from Supplier

    93
    ATCC f nucleatum atcc 10596
    F. nucleatum <t>ATCC</t> 23726 accelerates tumor growth and progression of lung metastasis. a Experimental scheme: AT3 breast cancer cell line was injected to the mammary fat pad of 6–7-week-old C57BL/6 mice. When tumor size reached 500 mm 3 , PBS vehicle (V), 5 × 10 7 F. nucleatum ATCC 23726 only (726), 5 × 10 7 F. nucleatum ATCC 23726 with metronidazole (726+MTZ.), metronidazole only (MTZ.), or 5 × 10 7 Fap2-deficient F. nucleatum K50 were injected into the tail vein. On days 2–3 post inoculation, mice were injected twice a day with metronidazole (M) or with PBS vehicle (V). On days 4–7 post inoculation, mice were injected once a day with metronidazole or with PBS. Eight days after inoculation, mice were sacrificed, and tumor and lungs harvested. Breast cancer tissues were weighed, and lung metastases were counted. b Tumor weights normalized as the percentage compared with the average tumor weight in the PBS group (vehicle control) in each experiment (set as 100% tumor weight). Each symbol represents one mouse. Presented results are of three independent experiments. **** p = 3.48 × 10 −5 , ** p = 0.002. Holm’s correction, one-tailed Mann–Whitney. Vehicle n = 21, MTZ. n = 17, K50 n = 7, 726 n = 19, 726+MTZ. n = 12. c Representative tumors post-harvest. d Representative image of lung metastases. Scale bar, 500 µm in the large image, 250 µm in the indicated inset. e Lung metastasis rank [number of metastases per lung area (pixel 2 )] calculated as percentage compared with the average metastasis rank in the vehicle control group of each experiment (set as 100% metastasis rank). Each symbol represents one mouse. Presented results are of three independent experiments. **** p = 4.08 × 10 −5 , * p = 0.0177, n.s.: non-significant, Holm’s correction, one-tailed Mann–Whitney. Vehicle n = 20, MTZ. n = 17, K50 n = 7, 726 n = 18, 726+met n = 12. f Images taken by fluorescence binocular of lung metastases in four mice implanted with GFP-expressing AT3 cells as described above. Two tumor-bearing mice were inoculated with F. nucleatum and two were treated by PBS vehicle. Scale bar, 1000 µm. Source data are provided as a Source data file.
    F Nucleatum Atcc 10596, supplied by ATCC, used in various techniques. Bioz Stars score: 93/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/f nucleatum atcc 10596/product/ATCC
    Average 93 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    f nucleatum atcc 10596 - by Bioz Stars, 2020-08
    93/100 stars
      Buy from Supplier

    92
    ATCC f nucleatum atcc 23276 competent cells
    F. nucleatum <t>ATCC</t> 23726 accelerates tumor growth and progression of lung metastasis. a Experimental scheme: AT3 breast cancer cell line was injected to the mammary fat pad of 6–7-week-old C57BL/6 mice. When tumor size reached 500 mm 3 , PBS vehicle (V), 5 × 10 7 F. nucleatum ATCC 23726 only (726), 5 × 10 7 F. nucleatum ATCC 23726 with metronidazole (726+MTZ.), metronidazole only (MTZ.), or 5 × 10 7 Fap2-deficient F. nucleatum K50 were injected into the tail vein. On days 2–3 post inoculation, mice were injected twice a day with metronidazole (M) or with PBS vehicle (V). On days 4–7 post inoculation, mice were injected once a day with metronidazole or with PBS. Eight days after inoculation, mice were sacrificed, and tumor and lungs harvested. Breast cancer tissues were weighed, and lung metastases were counted. b Tumor weights normalized as the percentage compared with the average tumor weight in the PBS group (vehicle control) in each experiment (set as 100% tumor weight). Each symbol represents one mouse. Presented results are of three independent experiments. **** p = 3.48 × 10 −5 , ** p = 0.002. Holm’s correction, one-tailed Mann–Whitney. Vehicle n = 21, MTZ. n = 17, K50 n = 7, 726 n = 19, 726+MTZ. n = 12. c Representative tumors post-harvest. d Representative image of lung metastases. Scale bar, 500 µm in the large image, 250 µm in the indicated inset. e Lung metastasis rank [number of metastases per lung area (pixel 2 )] calculated as percentage compared with the average metastasis rank in the vehicle control group of each experiment (set as 100% metastasis rank). Each symbol represents one mouse. Presented results are of three independent experiments. **** p = 4.08 × 10 −5 , * p = 0.0177, n.s.: non-significant, Holm’s correction, one-tailed Mann–Whitney. Vehicle n = 20, MTZ. n = 17, K50 n = 7, 726 n = 18, 726+met n = 12. f Images taken by fluorescence binocular of lung metastases in four mice implanted with GFP-expressing AT3 cells as described above. Two tumor-bearing mice were inoculated with F. nucleatum and two were treated by PBS vehicle. Scale bar, 1000 µm. Source data are provided as a Source data file.
    F Nucleatum Atcc 23276 Competent Cells, supplied by ATCC, used in various techniques. Bioz Stars score: 92/100, based on 14 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/f nucleatum atcc 23276 competent cells/product/ATCC
    Average 92 stars, based on 14 article reviews
    Price from $9.99 to $1999.99
    f nucleatum atcc 23276 competent cells - by Bioz Stars, 2020-08
    92/100 stars
      Buy from Supplier

    96
    ATCC f nucleatum atcc 2326
    F. nucleatum <t>ATCC</t> 23726 accelerates tumor growth and progression of lung metastasis. a Experimental scheme: AT3 breast cancer cell line was injected to the mammary fat pad of 6–7-week-old C57BL/6 mice. When tumor size reached 500 mm 3 , PBS vehicle (V), 5 × 10 7 F. nucleatum ATCC 23726 only (726), 5 × 10 7 F. nucleatum ATCC 23726 with metronidazole (726+MTZ.), metronidazole only (MTZ.), or 5 × 10 7 Fap2-deficient F. nucleatum K50 were injected into the tail vein. On days 2–3 post inoculation, mice were injected twice a day with metronidazole (M) or with PBS vehicle (V). On days 4–7 post inoculation, mice were injected once a day with metronidazole or with PBS. Eight days after inoculation, mice were sacrificed, and tumor and lungs harvested. Breast cancer tissues were weighed, and lung metastases were counted. b Tumor weights normalized as the percentage compared with the average tumor weight in the PBS group (vehicle control) in each experiment (set as 100% tumor weight). Each symbol represents one mouse. Presented results are of three independent experiments. **** p = 3.48 × 10 −5 , ** p = 0.002. Holm’s correction, one-tailed Mann–Whitney. Vehicle n = 21, MTZ. n = 17, K50 n = 7, 726 n = 19, 726+MTZ. n = 12. c Representative tumors post-harvest. d Representative image of lung metastases. Scale bar, 500 µm in the large image, 250 µm in the indicated inset. e Lung metastasis rank [number of metastases per lung area (pixel 2 )] calculated as percentage compared with the average metastasis rank in the vehicle control group of each experiment (set as 100% metastasis rank). Each symbol represents one mouse. Presented results are of three independent experiments. **** p = 4.08 × 10 −5 , * p = 0.0177, n.s.: non-significant, Holm’s correction, one-tailed Mann–Whitney. Vehicle n = 20, MTZ. n = 17, K50 n = 7, 726 n = 18, 726+met n = 12. f Images taken by fluorescence binocular of lung metastases in four mice implanted with GFP-expressing AT3 cells as described above. Two tumor-bearing mice were inoculated with F. nucleatum and two were treated by PBS vehicle. Scale bar, 1000 µm. Source data are provided as a Source data file.
    F Nucleatum Atcc 2326, supplied by ATCC, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/f nucleatum atcc 2326/product/ATCC
    Average 96 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    f nucleatum atcc 2326 - by Bioz Stars, 2020-08
    96/100 stars
      Buy from Supplier

    84
    ATCC bacterial culture f nucleatum atcc 25586
    F. nucleatum <t>ATCC</t> 23726 accelerates tumor growth and progression of lung metastasis. a Experimental scheme: AT3 breast cancer cell line was injected to the mammary fat pad of 6–7-week-old C57BL/6 mice. When tumor size reached 500 mm 3 , PBS vehicle (V), 5 × 10 7 F. nucleatum ATCC 23726 only (726), 5 × 10 7 F. nucleatum ATCC 23726 with metronidazole (726+MTZ.), metronidazole only (MTZ.), or 5 × 10 7 Fap2-deficient F. nucleatum K50 were injected into the tail vein. On days 2–3 post inoculation, mice were injected twice a day with metronidazole (M) or with PBS vehicle (V). On days 4–7 post inoculation, mice were injected once a day with metronidazole or with PBS. Eight days after inoculation, mice were sacrificed, and tumor and lungs harvested. Breast cancer tissues were weighed, and lung metastases were counted. b Tumor weights normalized as the percentage compared with the average tumor weight in the PBS group (vehicle control) in each experiment (set as 100% tumor weight). Each symbol represents one mouse. Presented results are of three independent experiments. **** p = 3.48 × 10 −5 , ** p = 0.002. Holm’s correction, one-tailed Mann–Whitney. Vehicle n = 21, MTZ. n = 17, K50 n = 7, 726 n = 19, 726+MTZ. n = 12. c Representative tumors post-harvest. d Representative image of lung metastases. Scale bar, 500 µm in the large image, 250 µm in the indicated inset. e Lung metastasis rank [number of metastases per lung area (pixel 2 )] calculated as percentage compared with the average metastasis rank in the vehicle control group of each experiment (set as 100% metastasis rank). Each symbol represents one mouse. Presented results are of three independent experiments. **** p = 4.08 × 10 −5 , * p = 0.0177, n.s.: non-significant, Holm’s correction, one-tailed Mann–Whitney. Vehicle n = 20, MTZ. n = 17, K50 n = 7, 726 n = 18, 726+met n = 12. f Images taken by fluorescence binocular of lung metastases in four mice implanted with GFP-expressing AT3 cells as described above. Two tumor-bearing mice were inoculated with F. nucleatum and two were treated by PBS vehicle. Scale bar, 1000 µm. Source data are provided as a Source data file.
    Bacterial Culture F Nucleatum Atcc 25586, supplied by ATCC, used in various techniques. Bioz Stars score: 84/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/bacterial culture f nucleatum atcc 25586/product/ATCC
    Average 84 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    bacterial culture f nucleatum atcc 25586 - by Bioz Stars, 2020-08
    84/100 stars
      Buy from Supplier

    84
    ATCC fap2 expressing f nucleatum atcc 23726
    F. nucleatum ATCC 23726 accelerates tumor growth and progression of lung metastasis. a Experimental scheme: AT3 breast cancer cell line was injected to the mammary fat pad of 6–7-week-old C57BL/6 mice. When tumor size reached 500 mm 3 , PBS vehicle (V), 5 × 10 7 F. nucleatum ATCC 23726 only (726), 5 × 10 7 F. nucleatum ATCC 23726 with metronidazole (726+MTZ.), metronidazole only (MTZ.), or 5 × 10 7 <t>Fap2-deficient</t> F. nucleatum K50 were injected into the tail vein. On days 2–3 post inoculation, mice were injected twice a day with metronidazole (M) or with PBS vehicle (V). On days 4–7 post inoculation, mice were injected once a day with metronidazole or with PBS. Eight days after inoculation, mice were sacrificed, and tumor and lungs harvested. Breast cancer tissues were weighed, and lung metastases were counted. b Tumor weights normalized as the percentage compared with the average tumor weight in the PBS group (vehicle control) in each experiment (set as 100% tumor weight). Each symbol represents one mouse. Presented results are of three independent experiments. **** p = 3.48 × 10 −5 , ** p = 0.002. Holm’s correction, one-tailed Mann–Whitney. Vehicle n = 21, MTZ. n = 17, K50 n = 7, 726 n = 19, 726+MTZ. n = 12. c Representative tumors post-harvest. d Representative image of lung metastases. Scale bar, 500 µm in the large image, 250 µm in the indicated inset. e Lung metastasis rank [number of metastases per lung area (pixel 2 )] calculated as percentage compared with the average metastasis rank in the vehicle control group of each experiment (set as 100% metastasis rank). Each symbol represents one mouse. Presented results are of three independent experiments. **** p = 4.08 × 10 −5 , * p = 0.0177, n.s.: non-significant, Holm’s correction, one-tailed Mann–Whitney. Vehicle n = 20, MTZ. n = 17, K50 n = 7, 726 n = 18, 726+met n = 12. f Images taken by fluorescence binocular of lung metastases in four mice implanted with GFP-expressing AT3 cells as described above. Two tumor-bearing mice were inoculated with F. nucleatum and two were treated by PBS vehicle. Scale bar, 1000 µm. Source data are provided as a Source data file.
    Fap2 Expressing F Nucleatum Atcc 23726, supplied by ATCC, used in various techniques. Bioz Stars score: 84/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/fap2 expressing f nucleatum atcc 23726/product/ATCC
    Average 84 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    fap2 expressing f nucleatum atcc 23726 - by Bioz Stars, 2020-08
    84/100 stars
      Buy from Supplier

    94
    ATCC f nucleatum atcc 49256
    Amino acid sequence alignment of FadA and its paralogues. Highlighted in gray are the identical residues shared among FadA proteins. The sequences of two FadA paralogues, FN1529 from F. <t>nucleatum</t> ATCC 25586 and FNV2159 from F. nucleatum ATCC 49256, are
    F Nucleatum Atcc 49256, supplied by ATCC, used in various techniques. Bioz Stars score: 94/100, based on 74 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/f nucleatum atcc 49256/product/ATCC
    Average 94 stars, based on 74 article reviews
    Price from $9.99 to $1999.99
    f nucleatum atcc 49256 - by Bioz Stars, 2020-08
    94/100 stars
      Buy from Supplier

    96
    ATCC f nucleatum atcc 25286
    Amino acid sequence alignment of FadA and its paralogues. Highlighted in gray are the identical residues shared among FadA proteins. The sequences of two FadA paralogues, FN1529 from F. <t>nucleatum</t> ATCC 25586 and FNV2159 from F. nucleatum ATCC 49256, are
    F Nucleatum Atcc 25286, supplied by ATCC, used in various techniques. Bioz Stars score: 96/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/f nucleatum atcc 25286/product/ATCC
    Average 96 stars, based on 3 article reviews
    Price from $9.99 to $1999.99
    f nucleatum atcc 25286 - by Bioz Stars, 2020-08
    96/100 stars
      Buy from Supplier

    84
    ATCC growth conditions f nucleatum strains atcc 25586
    AT3 mammary tumor acceleration by F. nucleatum is immune-mediated. a 1 × 10 6 AT3 cells were injected to the mammary fat pad of 6–7-week-old female C57BL/6 mice. When tumor volume reached 500 mm 3 , mice were IV injected with PBS vehicle or with 5 × 10 7 F. nucleatum ATCC 23726 (726). At day 8 post injection, mice were sacrificed, tumors were harvested subjected to flow cytometry and abundance of NK cells, CD4+ T cells and CD8+ T cells was determined. Boxplot whiskers represent extrema, box bounds represent upper and lower quartiles, and center-line represents the median value. n.s.: non-significant. ** p = 0.00216, * p = 0.0152, two-tailed Mann–Whitney (each symbol represents the mean of two technical replicates, n = 6 per group). b 1 × 10 5 AT3 or GFP-expressing AT3 cells were injected to the mammary fat pad of 6–7-week-old female SCID-beige mice. When tumor volume reached 500 mm 3 , mice were IV injected with PBS vehicle or with 5 × 10 7 F. nucleatum ATCC 23726 (726). At day 8 post injection, mice were sacrificed, tumors were harvested and weighed in an analytic weigh. Boxplot whiskers represent extrema, box bounds represent upper and lower quartiles, and center-line represents the median value of two independent experiments. n.s.: non-significant. n = 8 per vehicle group and n = 10 per F. nucleatum -infected group. c Representative picture and densitometry analysis of gelatin zymography of conditioned medium prepared from mouse mammary tumor cell line AT3 or from the human breast cancer cell line MDA-MB-231 that were co-cultured or not, with F. nucleatum strains <t>ATCC</t> 25586 or ATCC 23726. Bars indicate mean ± SEM of six independent experiments. * p = 0.031 two-tailed paired Wilcoxon. Note that mouse MMP-9 is larger than that of human. Source data are provided as a Source data file.
    Growth Conditions F Nucleatum Strains Atcc 25586, supplied by ATCC, used in various techniques. Bioz Stars score: 84/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/growth conditions f nucleatum strains atcc 25586/product/ATCC
    Average 84 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    growth conditions f nucleatum strains atcc 25586 - by Bioz Stars, 2020-08
    84/100 stars
      Buy from Supplier

    93
    atcc f nucleatum
    A model of F. <t>nucleatum</t> induced metastasis through cytokine signaling. We show that F. nucleatum induces IL-8 and CXCL1 secretion from CRC cells, and both cytokines have been characterized as key players in cancer metastasis and subsequent downstream cell seeding. As well as autocrine signaling back to cancer cells as a metastatic signal, HCT116 derived cytokines can participate in paracrine signaling to recruit neighboring immune cells, which further secrete their own cytokine signatures that alter the tumor microenvironment through metastatic, inflammatory, and immune cell programming.
    F Nucleatum, supplied by atcc, used in various techniques. Bioz Stars score: 93/100, based on 833 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/f nucleatum/product/atcc
    Average 93 stars, based on 833 article reviews
    Price from $9.99 to $1999.99
    f nucleatum - by Bioz Stars, 2020-08
    93/100 stars
      Buy from Supplier

    Image Search Results


    Effect of serum on viability of A. oris (Ao), V. parvula (Vp), F. nucleatum ATCC 10953 (Fn) and P. gingivalis 381 (Pg). Planktonic growth of microorganisms was evaluated in their appropriate nutrient-rich medium supplemented with 10% native human serum

    Journal: Anaerobe

    Article Title: Strain-Specific Colonization Patterns and Serum Modulation of Multi-Species Oral Biofilm Development

    doi: 10.1016/j.anaerobe.2012.06.003

    Figure Lengend Snippet: Effect of serum on viability of A. oris (Ao), V. parvula (Vp), F. nucleatum ATCC 10953 (Fn) and P. gingivalis 381 (Pg). Planktonic growth of microorganisms was evaluated in their appropriate nutrient-rich medium supplemented with 10% native human serum

    Article Snippet: For these experiments we first tested F. nucleatum ATCC 10953 since we previously identified a metabolic interaction occurring between this strain and P. gingivalis W50 [ ].

    Techniques:

    Planktonic growth of F. nucleatum ATCC 10953 in salivary growth medium as a mono-culture or in co-culture with A. oris and V. parvula . Panel A shows planktonic growth curves. Data points represent measurements from three independent replicate experiments.

    Journal: Anaerobe

    Article Title: Strain-Specific Colonization Patterns and Serum Modulation of Multi-Species Oral Biofilm Development

    doi: 10.1016/j.anaerobe.2012.06.003

    Figure Lengend Snippet: Planktonic growth of F. nucleatum ATCC 10953 in salivary growth medium as a mono-culture or in co-culture with A. oris and V. parvula . Panel A shows planktonic growth curves. Data points represent measurements from three independent replicate experiments.

    Article Snippet: For these experiments we first tested F. nucleatum ATCC 10953 since we previously identified a metabolic interaction occurring between this strain and P. gingivalis W50 [ ].

    Techniques: Co-Culture Assay

    The effect of the environmental conditions on the activity of H 2 S-producing enzymes was tested for Fusobacterium nucleatum polymorphum ATCC 10953. The bacteria were grown in different broths before protein extraction and 1D gel electrophoresis followed by in-gel activity assay

    Journal: BMC Microbiology

    Article Title: The proteins of Fusobacterium spp. involved in hydrogen sulfide production from L-cysteine

    doi: 10.1186/s12866-017-0967-9

    Figure Lengend Snippet: The effect of the environmental conditions on the activity of H 2 S-producing enzymes was tested for Fusobacterium nucleatum polymorphum ATCC 10953. The bacteria were grown in different broths before protein extraction and 1D gel electrophoresis followed by in-gel activity assay

    Article Snippet: In addition, we undertook to examine whether a cysteine-rich growth environment induced synthesis of H2 S producing enzymes and other intracellular enzymes in F. nucleatum polymorphum ATCC 10953.

    Techniques: Activity Assay, Protein Extraction, Nucleic Acid Electrophoresis

    PMSF inhibits the proteolytic activity of F. nucleatum . A, F. nucleatum FDC 364. B, F. nucleatum ATCC 25586. C, F. nucleatum 12230. D, F. nucleatum ATCC 23726. M, Molecular weight markers. Presented data are of representative zymograms.

    Journal: PLoS ONE

    Article Title: Identification and Characterization of Fusolisin, the Fusobacterium nucleatum Autotransporter Serine Protease

    doi: 10.1371/journal.pone.0111329

    Figure Lengend Snippet: PMSF inhibits the proteolytic activity of F. nucleatum . A, F. nucleatum FDC 364. B, F. nucleatum ATCC 25586. C, F. nucleatum 12230. D, F. nucleatum ATCC 23726. M, Molecular weight markers. Presented data are of representative zymograms.

    Article Snippet: The fact that Fsp25586 was not cleaved when expressed in F. nucleatum ATCC 23726 suggests that the processing of Fsp25586 is not efficient compared to that in Fsp23726 (and the orthologs in the other tested F. nucleatum strains, ).

    Techniques: Activity Assay, Molecular Weight

    Self-restriction of Fsp25586 is not efficient. Zymogram analysis of cell culture supernatant prepared from F. nucleatum ATCC 23726 carrying the pHS30 vector (A), or the pHSPROT plasmid expressing Fsp25586 (B).

    Journal: PLoS ONE

    Article Title: Identification and Characterization of Fusolisin, the Fusobacterium nucleatum Autotransporter Serine Protease

    doi: 10.1371/journal.pone.0111329

    Figure Lengend Snippet: Self-restriction of Fsp25586 is not efficient. Zymogram analysis of cell culture supernatant prepared from F. nucleatum ATCC 23726 carrying the pHS30 vector (A), or the pHSPROT plasmid expressing Fsp25586 (B).

    Article Snippet: The fact that Fsp25586 was not cleaved when expressed in F. nucleatum ATCC 23726 suggests that the processing of Fsp25586 is not efficient compared to that in Fsp23726 (and the orthologs in the other tested F. nucleatum strains, ).

    Techniques: Cell Culture, Plasmid Preparation, Expressing

    Protease profiles of F. nucleatum growth medium supernatants on fibrinogen containing zymograms. M, Molecular weight markers. A, F. nucleatum ATCC 49256. B, F. nucleatum FDC 364. C, F. nucleatum ATCC 10953. D, F. nucleatum ATCC 25586. E, F. nucleatum ATCC 23726. F, F. nucleatum 12230. Arrows indicate proteolytic bands. Presented data are of representative zymograms.

    Journal: PLoS ONE

    Article Title: Identification and Characterization of Fusolisin, the Fusobacterium nucleatum Autotransporter Serine Protease

    doi: 10.1371/journal.pone.0111329

    Figure Lengend Snippet: Protease profiles of F. nucleatum growth medium supernatants on fibrinogen containing zymograms. M, Molecular weight markers. A, F. nucleatum ATCC 49256. B, F. nucleatum FDC 364. C, F. nucleatum ATCC 10953. D, F. nucleatum ATCC 25586. E, F. nucleatum ATCC 23726. F, F. nucleatum 12230. Arrows indicate proteolytic bands. Presented data are of representative zymograms.

    Article Snippet: The fact that Fsp25586 was not cleaved when expressed in F. nucleatum ATCC 23726 suggests that the processing of Fsp25586 is not efficient compared to that in Fsp23726 (and the orthologs in the other tested F. nucleatum strains, ).

    Techniques: Molecular Weight

    pKH9 axf-txf toxin-antitoxin system. (A) Stability of a pKH9 derivative containing axf-txf (pKH90) (triangles) versus that of a derivative lacking axf-txf (pORI9) (squares) in F. nucleatum ATCC 23726. Open symbols represent results of the first experiment, and filled symbols represent those of a second independent experiment. (B) Region of overlapping sequence between the axf and txf genes.

    Journal: Applied and Environmental Microbiology

    Article Title: Characterization of the Novel Fusobacterium nucleatum Plasmid pKH9 and Evidence of an Addiction System

    doi: 10.1128/AEM.70.12.6957-6962.2004

    Figure Lengend Snippet: pKH9 axf-txf toxin-antitoxin system. (A) Stability of a pKH9 derivative containing axf-txf (pKH90) (triangles) versus that of a derivative lacking axf-txf (pORI9) (squares) in F. nucleatum ATCC 23726. Open symbols represent results of the first experiment, and filled symbols represent those of a second independent experiment. (B) Region of overlapping sequence between the axf and txf genes.

    Article Snippet: The 4,584-bp fragment released by digestion of pKH9 with XbaI was PCR amplified and cloned into the XbaI site of pBK-erm (Fig. ; Table ) to create pKH90, which confers clindamycin resistance on F. nucleatum ATCC 23726 transformants isolated after electroporation (Fig. ).

    Techniques: Sequencing

    Compatibility of pHS30 and pORI91. F. nucleatum ATCC 23726 harboring pHS30 was transformed with pORI91. Transformants were selected on media with clindamycin. Plasmid DNA isolated from representative transformants (T-1 and T-2) was examined by restriction enzyme digestion and compared with similarly digested pHS30 and pORI91 isolated from E. coli . DNA in lanes 1 was digested with KpnI to linearize pORI91. DNA in lanes 2 was digested with EcoRV to linearize pHS30. The open circular (OC), linear (L), and covalently closed circular (CCC) forms of pHS30 and pORI91 are indicated on the left and right, respectively.

    Journal: Applied and Environmental Microbiology

    Article Title: Characterization of the Novel Fusobacterium nucleatum Plasmid pKH9 and Evidence of an Addiction System

    doi: 10.1128/AEM.70.12.6957-6962.2004

    Figure Lengend Snippet: Compatibility of pHS30 and pORI91. F. nucleatum ATCC 23726 harboring pHS30 was transformed with pORI91. Transformants were selected on media with clindamycin. Plasmid DNA isolated from representative transformants (T-1 and T-2) was examined by restriction enzyme digestion and compared with similarly digested pHS30 and pORI91 isolated from E. coli . DNA in lanes 1 was digested with KpnI to linearize pORI91. DNA in lanes 2 was digested with EcoRV to linearize pHS30. The open circular (OC), linear (L), and covalently closed circular (CCC) forms of pHS30 and pORI91 are indicated on the left and right, respectively.

    Article Snippet: The 4,584-bp fragment released by digestion of pKH9 with XbaI was PCR amplified and cloned into the XbaI site of pBK-erm (Fig. ; Table ) to create pKH90, which confers clindamycin resistance on F. nucleatum ATCC 23726 transformants isolated after electroporation (Fig. ).

    Techniques: Transformation Assay, Plasmid Preparation, Isolation, Countercurrent Chromatography

    (a and b) RANKL expression in PDL cells stimulated by F. nucleatum ATCC 25586 and/or CTSH at 1 day (a) and 3 days (b). (c and d) OPG expression in PDL cells stimulated by F. nucleatum ATCC 25586 and/or CTSH at 1 day (c) and 3 days (d). (e and f) RANKL/OPG mRNA ratio in PDL cells stimulated by F. nucleatum ATCC 25586 and/or CTSH at 1 day (e) and 3 days (f). *Significant difference compared to other groups ( P

    Journal: Mediators of Inflammation

    Article Title: Biomechanical Loading Modulates Proinflammatory and Bone Resorptive Mediators in Bacterial-Stimulated PDL Cells

    doi: 10.1155/2014/425421

    Figure Lengend Snippet: (a and b) RANKL expression in PDL cells stimulated by F. nucleatum ATCC 25586 and/or CTSH at 1 day (a) and 3 days (b). (c and d) OPG expression in PDL cells stimulated by F. nucleatum ATCC 25586 and/or CTSH at 1 day (c) and 3 days (d). (e and f) RANKL/OPG mRNA ratio in PDL cells stimulated by F. nucleatum ATCC 25586 and/or CTSH at 1 day (e) and 3 days (f). *Significant difference compared to other groups ( P

    Article Snippet: The association of CTSH and F. nucleatum ATCC 25586 resulted in an RANKL/OPG ratio that is significantly higher when compared to F. nucleatum ATCC 25586 alone.

    Techniques: Expressing

    (a and b) Synthesis of RANKL protein in supernatants of PDL cells stimulated by F. nucleatum ATCC 25586 and/or CTSH at 1 day (a) and 3 days (b). (c and d) Synthesis of OPG protein in supernatants of PDL cells stimulated by F. nucleatum ATCC 25586 and/or CTSH at 1 day (c) and 3 days (d). (e and f) RANKL/OPG protein ratio in supernatants of PDL cells stimulated by F. nucleatum ATCC 25586 and/or CTSH at 1 day (e) and 3 days (f). *Significant difference compared to other groups ( P

    Journal: Mediators of Inflammation

    Article Title: Biomechanical Loading Modulates Proinflammatory and Bone Resorptive Mediators in Bacterial-Stimulated PDL Cells

    doi: 10.1155/2014/425421

    Figure Lengend Snippet: (a and b) Synthesis of RANKL protein in supernatants of PDL cells stimulated by F. nucleatum ATCC 25586 and/or CTSH at 1 day (a) and 3 days (b). (c and d) Synthesis of OPG protein in supernatants of PDL cells stimulated by F. nucleatum ATCC 25586 and/or CTSH at 1 day (c) and 3 days (d). (e and f) RANKL/OPG protein ratio in supernatants of PDL cells stimulated by F. nucleatum ATCC 25586 and/or CTSH at 1 day (e) and 3 days (f). *Significant difference compared to other groups ( P

    Article Snippet: The association of CTSH and F. nucleatum ATCC 25586 resulted in an RANKL/OPG ratio that is significantly higher when compared to F. nucleatum ATCC 25586 alone.

    Techniques:

    (a) COX2 expression in PDL cells stimulated by F. nucleatum ATCC 25586 over time. (b) COX2 expression in PDL cells stimulated by various concentrations of F. nucleatum ATCC 25586 at 1 day. (c) OPG expression in PDL cells stimulated by F. nucleatum ATCC 25586 over time. (d) OPG expression in PDL cells stimulated by various concentrations of F. nucleatum ATCC 25586 at 1 day. (e) COX2 expression in PDL cells stimulated by F. nucleatum ATCC 25586 (OD 0.025) in the presence or in the absence of anti-TLR2 or anti-TLR4 antibodies at 1 day. *Significant difference between groups ( P

    Journal: Mediators of Inflammation

    Article Title: Biomechanical Loading Modulates Proinflammatory and Bone Resorptive Mediators in Bacterial-Stimulated PDL Cells

    doi: 10.1155/2014/425421

    Figure Lengend Snippet: (a) COX2 expression in PDL cells stimulated by F. nucleatum ATCC 25586 over time. (b) COX2 expression in PDL cells stimulated by various concentrations of F. nucleatum ATCC 25586 at 1 day. (c) OPG expression in PDL cells stimulated by F. nucleatum ATCC 25586 over time. (d) OPG expression in PDL cells stimulated by various concentrations of F. nucleatum ATCC 25586 at 1 day. (e) COX2 expression in PDL cells stimulated by F. nucleatum ATCC 25586 (OD 0.025) in the presence or in the absence of anti-TLR2 or anti-TLR4 antibodies at 1 day. *Significant difference between groups ( P

    Article Snippet: The association of CTSH and F. nucleatum ATCC 25586 resulted in an RANKL/OPG ratio that is significantly higher when compared to F. nucleatum ATCC 25586 alone.

    Techniques: Expressing

    (a) Synthesis of COX2 in lysates of PDL cells treated with F. nucleatum ATCC 25586 and/or cyclic tensile strain (CTS) of low (CTSL, 3%) and high (CTSH, 20%) magnitudes at 36 hours. (b and c) Production of PGE2 in supernatants of PDL cells treated with F. nucleatum ATCC 25586 and/or CTSL and CTSH at 1 day (b) and 3 days (c). *Significant difference compared to other groups ( P

    Journal: Mediators of Inflammation

    Article Title: Biomechanical Loading Modulates Proinflammatory and Bone Resorptive Mediators in Bacterial-Stimulated PDL Cells

    doi: 10.1155/2014/425421

    Figure Lengend Snippet: (a) Synthesis of COX2 in lysates of PDL cells treated with F. nucleatum ATCC 25586 and/or cyclic tensile strain (CTS) of low (CTSL, 3%) and high (CTSH, 20%) magnitudes at 36 hours. (b and c) Production of PGE2 in supernatants of PDL cells treated with F. nucleatum ATCC 25586 and/or CTSL and CTSH at 1 day (b) and 3 days (c). *Significant difference compared to other groups ( P

    Article Snippet: The association of CTSH and F. nucleatum ATCC 25586 resulted in an RANKL/OPG ratio that is significantly higher when compared to F. nucleatum ATCC 25586 alone.

    Techniques:

    (A) Western immunoblot of FAD-I protein in whole-cell lysates of F. nucleatum ATCC 25586, ATCC 23726, ATCC 10953, ATCC 10953 Δ fadI , and ATCC 10953 Δ fadI /pFAD-I 23726 (ATCC 10953 Δ fadI with plasmid-based expression of FAD-I from

    Journal: Infection and Immunity

    Article Title: FAD-I, a Fusobacterium nucleatum Cell Wall-Associated Diacylated Lipoprotein That Mediates Human Beta Defensin 2 Induction through Toll-Like Receptor-1/2 (TLR-1/2) and TLR-2/6

    doi: 10.1128/IAI.01311-15

    Figure Lengend Snippet: (A) Western immunoblot of FAD-I protein in whole-cell lysates of F. nucleatum ATCC 25586, ATCC 23726, ATCC 10953, ATCC 10953 Δ fadI , and ATCC 10953 Δ fadI /pFAD-I 23726 (ATCC 10953 Δ fadI with plasmid-based expression of FAD-I from

    Article Snippet: The full-length FAD-I gene (FN1527) ( ) was amplified by PCR from genomic DNA of F. nucleatum strain ATCC 25586 using primers (see Table S2 in the supplemental material) with Nde1 and Xho1 restriction sites at their ends (New England BioLabs).

    Techniques: Western Blot, Plasmid Preparation, Expressing

    Induction of hBD-2 mRNA by live bacterial cells (A) and cell wall preparations (10 μg/ml cell wall F. nucleatum [Fn] ATCC 25586, ATCC 23726, and ATCC 10953 (B). The data presented are means ± standard deviations (SD) of the results of

    Journal: Infection and Immunity

    Article Title: FAD-I, a Fusobacterium nucleatum Cell Wall-Associated Diacylated Lipoprotein That Mediates Human Beta Defensin 2 Induction through Toll-Like Receptor-1/2 (TLR-1/2) and TLR-2/6

    doi: 10.1128/IAI.01311-15

    Figure Lengend Snippet: Induction of hBD-2 mRNA by live bacterial cells (A) and cell wall preparations (10 μg/ml cell wall F. nucleatum [Fn] ATCC 25586, ATCC 23726, and ATCC 10953 (B). The data presented are means ± standard deviations (SD) of the results of

    Article Snippet: The full-length FAD-I gene (FN1527) ( ) was amplified by PCR from genomic DNA of F. nucleatum strain ATCC 25586 using primers (see Table S2 in the supplemental material) with Nde1 and Xho1 restriction sites at their ends (New England BioLabs).

    Techniques:

    F. nucleatum ATCC 23726 accelerates tumor growth and progression of lung metastasis. a Experimental scheme: AT3 breast cancer cell line was injected to the mammary fat pad of 6–7-week-old C57BL/6 mice. When tumor size reached 500 mm 3 , PBS vehicle (V), 5 × 10 7 F. nucleatum ATCC 23726 only (726), 5 × 10 7 F. nucleatum ATCC 23726 with metronidazole (726+MTZ.), metronidazole only (MTZ.), or 5 × 10 7 Fap2-deficient F. nucleatum K50 were injected into the tail vein. On days 2–3 post inoculation, mice were injected twice a day with metronidazole (M) or with PBS vehicle (V). On days 4–7 post inoculation, mice were injected once a day with metronidazole or with PBS. Eight days after inoculation, mice were sacrificed, and tumor and lungs harvested. Breast cancer tissues were weighed, and lung metastases were counted. b Tumor weights normalized as the percentage compared with the average tumor weight in the PBS group (vehicle control) in each experiment (set as 100% tumor weight). Each symbol represents one mouse. Presented results are of three independent experiments. **** p = 3.48 × 10 −5 , ** p = 0.002. Holm’s correction, one-tailed Mann–Whitney. Vehicle n = 21, MTZ. n = 17, K50 n = 7, 726 n = 19, 726+MTZ. n = 12. c Representative tumors post-harvest. d Representative image of lung metastases. Scale bar, 500 µm in the large image, 250 µm in the indicated inset. e Lung metastasis rank [number of metastases per lung area (pixel 2 )] calculated as percentage compared with the average metastasis rank in the vehicle control group of each experiment (set as 100% metastasis rank). Each symbol represents one mouse. Presented results are of three independent experiments. **** p = 4.08 × 10 −5 , * p = 0.0177, n.s.: non-significant, Holm’s correction, one-tailed Mann–Whitney. Vehicle n = 20, MTZ. n = 17, K50 n = 7, 726 n = 18, 726+met n = 12. f Images taken by fluorescence binocular of lung metastases in four mice implanted with GFP-expressing AT3 cells as described above. Two tumor-bearing mice were inoculated with F. nucleatum and two were treated by PBS vehicle. Scale bar, 1000 µm. Source data are provided as a Source data file.

    Journal: Nature Communications

    Article Title: Breast cancer colonization by Fusobacterium nucleatum accelerates tumor growth and metastatic progression

    doi: 10.1038/s41467-020-16967-2

    Figure Lengend Snippet: F. nucleatum ATCC 23726 accelerates tumor growth and progression of lung metastasis. a Experimental scheme: AT3 breast cancer cell line was injected to the mammary fat pad of 6–7-week-old C57BL/6 mice. When tumor size reached 500 mm 3 , PBS vehicle (V), 5 × 10 7 F. nucleatum ATCC 23726 only (726), 5 × 10 7 F. nucleatum ATCC 23726 with metronidazole (726+MTZ.), metronidazole only (MTZ.), or 5 × 10 7 Fap2-deficient F. nucleatum K50 were injected into the tail vein. On days 2–3 post inoculation, mice were injected twice a day with metronidazole (M) or with PBS vehicle (V). On days 4–7 post inoculation, mice were injected once a day with metronidazole or with PBS. Eight days after inoculation, mice were sacrificed, and tumor and lungs harvested. Breast cancer tissues were weighed, and lung metastases were counted. b Tumor weights normalized as the percentage compared with the average tumor weight in the PBS group (vehicle control) in each experiment (set as 100% tumor weight). Each symbol represents one mouse. Presented results are of three independent experiments. **** p = 3.48 × 10 −5 , ** p = 0.002. Holm’s correction, one-tailed Mann–Whitney. Vehicle n = 21, MTZ. n = 17, K50 n = 7, 726 n = 19, 726+MTZ. n = 12. c Representative tumors post-harvest. d Representative image of lung metastases. Scale bar, 500 µm in the large image, 250 µm in the indicated inset. e Lung metastasis rank [number of metastases per lung area (pixel 2 )] calculated as percentage compared with the average metastasis rank in the vehicle control group of each experiment (set as 100% metastasis rank). Each symbol represents one mouse. Presented results are of three independent experiments. **** p = 4.08 × 10 −5 , * p = 0.0177, n.s.: non-significant, Holm’s correction, one-tailed Mann–Whitney. Vehicle n = 20, MTZ. n = 17, K50 n = 7, 726 n = 18, 726+met n = 12. f Images taken by fluorescence binocular of lung metastases in four mice implanted with GFP-expressing AT3 cells as described above. Two tumor-bearing mice were inoculated with F. nucleatum and two were treated by PBS vehicle. Scale bar, 1000 µm. Source data are provided as a Source data file.

    Article Snippet: Supporting the predicted Fap2-mediated recognition of Gal-GalNAc, wild-type F. nucleatum ATCC 23726 showed significantly higher attachment than the K50 and D22 mutants (Fig. ).

    Techniques: Injection, Mouse Assay, One-tailed Test, MANN-WHITNEY, Fluorescence, Expressing

    AT3 mammary tumor acceleration by F. nucleatum is immune-mediated. a 1 × 10 6 AT3 cells were injected to the mammary fat pad of 6–7-week-old female C57BL/6 mice. When tumor volume reached 500 mm 3 , mice were IV injected with PBS vehicle or with 5 × 10 7 F. nucleatum ATCC 23726 (726). At day 8 post injection, mice were sacrificed, tumors were harvested subjected to flow cytometry and abundance of NK cells, CD4+ T cells and CD8+ T cells was determined. Boxplot whiskers represent extrema, box bounds represent upper and lower quartiles, and center-line represents the median value. n.s.: non-significant. ** p = 0.00216, * p = 0.0152, two-tailed Mann–Whitney (each symbol represents the mean of two technical replicates, n = 6 per group). b 1 × 10 5 AT3 or GFP-expressing AT3 cells were injected to the mammary fat pad of 6–7-week-old female SCID-beige mice. When tumor volume reached 500 mm 3 , mice were IV injected with PBS vehicle or with 5 × 10 7 F. nucleatum ATCC 23726 (726). At day 8 post injection, mice were sacrificed, tumors were harvested and weighed in an analytic weigh. Boxplot whiskers represent extrema, box bounds represent upper and lower quartiles, and center-line represents the median value of two independent experiments. n.s.: non-significant. n = 8 per vehicle group and n = 10 per F. nucleatum -infected group. c Representative picture and densitometry analysis of gelatin zymography of conditioned medium prepared from mouse mammary tumor cell line AT3 or from the human breast cancer cell line MDA-MB-231 that were co-cultured or not, with F. nucleatum strains ATCC 25586 or ATCC 23726. Bars indicate mean ± SEM of six independent experiments. * p = 0.031 two-tailed paired Wilcoxon. Note that mouse MMP-9 is larger than that of human. Source data are provided as a Source data file.

    Journal: Nature Communications

    Article Title: Breast cancer colonization by Fusobacterium nucleatum accelerates tumor growth and metastatic progression

    doi: 10.1038/s41467-020-16967-2

    Figure Lengend Snippet: AT3 mammary tumor acceleration by F. nucleatum is immune-mediated. a 1 × 10 6 AT3 cells were injected to the mammary fat pad of 6–7-week-old female C57BL/6 mice. When tumor volume reached 500 mm 3 , mice were IV injected with PBS vehicle or with 5 × 10 7 F. nucleatum ATCC 23726 (726). At day 8 post injection, mice were sacrificed, tumors were harvested subjected to flow cytometry and abundance of NK cells, CD4+ T cells and CD8+ T cells was determined. Boxplot whiskers represent extrema, box bounds represent upper and lower quartiles, and center-line represents the median value. n.s.: non-significant. ** p = 0.00216, * p = 0.0152, two-tailed Mann–Whitney (each symbol represents the mean of two technical replicates, n = 6 per group). b 1 × 10 5 AT3 or GFP-expressing AT3 cells were injected to the mammary fat pad of 6–7-week-old female SCID-beige mice. When tumor volume reached 500 mm 3 , mice were IV injected with PBS vehicle or with 5 × 10 7 F. nucleatum ATCC 23726 (726). At day 8 post injection, mice were sacrificed, tumors were harvested and weighed in an analytic weigh. Boxplot whiskers represent extrema, box bounds represent upper and lower quartiles, and center-line represents the median value of two independent experiments. n.s.: non-significant. n = 8 per vehicle group and n = 10 per F. nucleatum -infected group. c Representative picture and densitometry analysis of gelatin zymography of conditioned medium prepared from mouse mammary tumor cell line AT3 or from the human breast cancer cell line MDA-MB-231 that were co-cultured or not, with F. nucleatum strains ATCC 25586 or ATCC 23726. Bars indicate mean ± SEM of six independent experiments. * p = 0.031 two-tailed paired Wilcoxon. Note that mouse MMP-9 is larger than that of human. Source data are provided as a Source data file.

    Article Snippet: Supporting the predicted Fap2-mediated recognition of Gal-GalNAc, wild-type F. nucleatum ATCC 23726 showed significantly higher attachment than the K50 and D22 mutants (Fig. ).

    Techniques: Injection, Mouse Assay, Flow Cytometry, Two Tailed Test, MANN-WHITNEY, Expressing, Infection, Zymography, Multiple Displacement Amplification, Cell Culture

    F. nucleatum colonizes mammary cancer in a Fap2-mediated mechanism. a Orthotropic 4T1 BALB/c mouse mammary cancer model. When reaching 500 mm 3 size, 5 × 10 7 F. nucleatum ATCC 23726 were IV injected. Twenty-four hours post inoculation, breast cancer and normal tissue from an adjacent mammary were harvested. b Representative images and sum of fluorescence intensity of binding of FITC-labeled PNA (green) to tumor and normal tissue stained with Hoechst dye (blue). Each paired symbols represents one mouse ( n = 7), ** p = 0.0078, one-tailed, Wilcoxon matched-paired signed rank test. c Relative F. nucleatum gDNA abundance (2 −ΔCt ) in breast tissue from tumor-free mice (No tumor, n = 3), and in tumor and matching normal tissue of 4T1 tumor-bearing mice ( n = 5) all inoculated with F. nucleatum ATCC 23726. * p = 0.0358 Holm-corrected one-tailed Mann–Whitney test. * p = 0.0313, one-tailed, Wilcoxon matched-paired signed rank test (for paired normal adjacent-tumor samples). d Orthotropic AT3 C57BL/6 mouse mammary cancer model. When reaching 500 mm 3 , mice were inoculated with 5 × 10 7 F. nucleatum ATCC 23726 (726; blue), 5 × 10 7 Fap2 mutant K50 (MUT K50; red) or with 5 × 10 7 P. gingivalis (Pg; black). Twenty-four hours later, normal mammary and breast cancer tissue were harvested from each mice and bacteria quantified. e Bacterial abundance (CFU/g tissue), and relative bacterial gDNA abundance (2 −ΔCt ) in tumor and normal tissue of AT3 tumor-bearing mice inoculated with P. gingivalis ( Pg , n = 9), K50 ( n = 12), or F. nucleatum ATCC 23726 (726, n = 12). Each symbol represents one mouse. * p = 0.0156 one-tailed, Wilcoxon matched-paired signed rank test. *** p = 0.0006, ** p = 0.001598. Holm-corrected one-tailed Mann–Whitney test (left panel). **** p

    Journal: Nature Communications

    Article Title: Breast cancer colonization by Fusobacterium nucleatum accelerates tumor growth and metastatic progression

    doi: 10.1038/s41467-020-16967-2

    Figure Lengend Snippet: F. nucleatum colonizes mammary cancer in a Fap2-mediated mechanism. a Orthotropic 4T1 BALB/c mouse mammary cancer model. When reaching 500 mm 3 size, 5 × 10 7 F. nucleatum ATCC 23726 were IV injected. Twenty-four hours post inoculation, breast cancer and normal tissue from an adjacent mammary were harvested. b Representative images and sum of fluorescence intensity of binding of FITC-labeled PNA (green) to tumor and normal tissue stained with Hoechst dye (blue). Each paired symbols represents one mouse ( n = 7), ** p = 0.0078, one-tailed, Wilcoxon matched-paired signed rank test. c Relative F. nucleatum gDNA abundance (2 −ΔCt ) in breast tissue from tumor-free mice (No tumor, n = 3), and in tumor and matching normal tissue of 4T1 tumor-bearing mice ( n = 5) all inoculated with F. nucleatum ATCC 23726. * p = 0.0358 Holm-corrected one-tailed Mann–Whitney test. * p = 0.0313, one-tailed, Wilcoxon matched-paired signed rank test (for paired normal adjacent-tumor samples). d Orthotropic AT3 C57BL/6 mouse mammary cancer model. When reaching 500 mm 3 , mice were inoculated with 5 × 10 7 F. nucleatum ATCC 23726 (726; blue), 5 × 10 7 Fap2 mutant K50 (MUT K50; red) or with 5 × 10 7 P. gingivalis (Pg; black). Twenty-four hours later, normal mammary and breast cancer tissue were harvested from each mice and bacteria quantified. e Bacterial abundance (CFU/g tissue), and relative bacterial gDNA abundance (2 −ΔCt ) in tumor and normal tissue of AT3 tumor-bearing mice inoculated with P. gingivalis ( Pg , n = 9), K50 ( n = 12), or F. nucleatum ATCC 23726 (726, n = 12). Each symbol represents one mouse. * p = 0.0156 one-tailed, Wilcoxon matched-paired signed rank test. *** p = 0.0006, ** p = 0.001598. Holm-corrected one-tailed Mann–Whitney test (left panel). **** p

    Article Snippet: Supporting the predicted Fap2-mediated recognition of Gal-GalNAc, wild-type F. nucleatum ATCC 23726 showed significantly higher attachment than the K50 and D22 mutants (Fig. ).

    Techniques: Injection, Fluorescence, Binding Assay, Labeling, Staining, One-tailed Test, Mouse Assay, MANN-WHITNEY, Mutagenesis

    Attachment of F. nucleatum ATCC 23726 to breast cancer is Fap2-mediated and Gal-GalNAc dependent. a Representative images (left panels, scale bar 20 µm) and quantitative analysis (Fn/mm 2 ) (right panel) of binding of Cy3-labeled (white) WT F. nucleatum ATCC 23726 (726) and of its Cy5-labeled (red) Fap2-inactivated isogenic mutant K50, to Hoechst-stained (blue) human breast cancer TMAs (HBre-Duc060CS-01 and BR1006). Results in the right panel are classified by tissue type (normal n = 35, benign n = 7, malignant n = 64) and bacterial strain (K50, 726). Each core is measured twice, once for each of the strains in the appropriate tissue type (as demonstrated in the pictures). When an individual contributed a single core to the experiment, the two observations are shown as circles (●); when she contributed two cores, these were to the normal type and the malignant type, and the four observations are depicted as plus signs (+). The six classes were compared against each other in pairs: the exact two-tailed Wilcoxon test was used for pairs that were from the same core, the exact two-tailed Mann–Whitney test for independent samples. Where the same hypothesis was tested on more than one pair, and the individual pairs were mutually independent, the corresponding p -values were combined by Fisher’s method. Wherever the pairs were not independent, the multiple comparisons were adjusted for using the Holm modification of the Bonferroni correction. **** p

    Journal: Nature Communications

    Article Title: Breast cancer colonization by Fusobacterium nucleatum accelerates tumor growth and metastatic progression

    doi: 10.1038/s41467-020-16967-2

    Figure Lengend Snippet: Attachment of F. nucleatum ATCC 23726 to breast cancer is Fap2-mediated and Gal-GalNAc dependent. a Representative images (left panels, scale bar 20 µm) and quantitative analysis (Fn/mm 2 ) (right panel) of binding of Cy3-labeled (white) WT F. nucleatum ATCC 23726 (726) and of its Cy5-labeled (red) Fap2-inactivated isogenic mutant K50, to Hoechst-stained (blue) human breast cancer TMAs (HBre-Duc060CS-01 and BR1006). Results in the right panel are classified by tissue type (normal n = 35, benign n = 7, malignant n = 64) and bacterial strain (K50, 726). Each core is measured twice, once for each of the strains in the appropriate tissue type (as demonstrated in the pictures). When an individual contributed a single core to the experiment, the two observations are shown as circles (●); when she contributed two cores, these were to the normal type and the malignant type, and the four observations are depicted as plus signs (+). The six classes were compared against each other in pairs: the exact two-tailed Wilcoxon test was used for pairs that were from the same core, the exact two-tailed Mann–Whitney test for independent samples. Where the same hypothesis was tested on more than one pair, and the individual pairs were mutually independent, the corresponding p -values were combined by Fisher’s method. Wherever the pairs were not independent, the multiple comparisons were adjusted for using the Holm modification of the Bonferroni correction. **** p

    Article Snippet: Supporting the predicted Fap2-mediated recognition of Gal-GalNAc, wild-type F. nucleatum ATCC 23726 showed significantly higher attachment than the K50 and D22 mutants (Fig. ).

    Techniques: Binding Assay, Labeling, Mutagenesis, Staining, Two Tailed Test, MANN-WHITNEY, Modification

    F. nucleatum ATCC 23726 accelerates tumor growth and progression of lung metastasis. a Experimental scheme: AT3 breast cancer cell line was injected to the mammary fat pad of 6–7-week-old C57BL/6 mice. When tumor size reached 500 mm 3 , PBS vehicle (V), 5 × 10 7 F. nucleatum ATCC 23726 only (726), 5 × 10 7 F. nucleatum ATCC 23726 with metronidazole (726+MTZ.), metronidazole only (MTZ.), or 5 × 10 7 Fap2-deficient F. nucleatum K50 were injected into the tail vein. On days 2–3 post inoculation, mice were injected twice a day with metronidazole (M) or with PBS vehicle (V). On days 4–7 post inoculation, mice were injected once a day with metronidazole or with PBS. Eight days after inoculation, mice were sacrificed, and tumor and lungs harvested. Breast cancer tissues were weighed, and lung metastases were counted. b Tumor weights normalized as the percentage compared with the average tumor weight in the PBS group (vehicle control) in each experiment (set as 100% tumor weight). Each symbol represents one mouse. Presented results are of three independent experiments. **** p = 3.48 × 10 −5 , ** p = 0.002. Holm’s correction, one-tailed Mann–Whitney. Vehicle n = 21, MTZ. n = 17, K50 n = 7, 726 n = 19, 726+MTZ. n = 12. c Representative tumors post-harvest. d Representative image of lung metastases. Scale bar, 500 µm in the large image, 250 µm in the indicated inset. e Lung metastasis rank [number of metastases per lung area (pixel 2 )] calculated as percentage compared with the average metastasis rank in the vehicle control group of each experiment (set as 100% metastasis rank). Each symbol represents one mouse. Presented results are of three independent experiments. **** p = 4.08 × 10 −5 , * p = 0.0177, n.s.: non-significant, Holm’s correction, one-tailed Mann–Whitney. Vehicle n = 20, MTZ. n = 17, K50 n = 7, 726 n = 18, 726+met n = 12. f Images taken by fluorescence binocular of lung metastases in four mice implanted with GFP-expressing AT3 cells as described above. Two tumor-bearing mice were inoculated with F. nucleatum and two were treated by PBS vehicle. Scale bar, 1000 µm. Source data are provided as a Source data file.

    Journal: Nature Communications

    Article Title: Breast cancer colonization by Fusobacterium nucleatum accelerates tumor growth and metastatic progression

    doi: 10.1038/s41467-020-16967-2

    Figure Lengend Snippet: F. nucleatum ATCC 23726 accelerates tumor growth and progression of lung metastasis. a Experimental scheme: AT3 breast cancer cell line was injected to the mammary fat pad of 6–7-week-old C57BL/6 mice. When tumor size reached 500 mm 3 , PBS vehicle (V), 5 × 10 7 F. nucleatum ATCC 23726 only (726), 5 × 10 7 F. nucleatum ATCC 23726 with metronidazole (726+MTZ.), metronidazole only (MTZ.), or 5 × 10 7 Fap2-deficient F. nucleatum K50 were injected into the tail vein. On days 2–3 post inoculation, mice were injected twice a day with metronidazole (M) or with PBS vehicle (V). On days 4–7 post inoculation, mice were injected once a day with metronidazole or with PBS. Eight days after inoculation, mice were sacrificed, and tumor and lungs harvested. Breast cancer tissues were weighed, and lung metastases were counted. b Tumor weights normalized as the percentage compared with the average tumor weight in the PBS group (vehicle control) in each experiment (set as 100% tumor weight). Each symbol represents one mouse. Presented results are of three independent experiments. **** p = 3.48 × 10 −5 , ** p = 0.002. Holm’s correction, one-tailed Mann–Whitney. Vehicle n = 21, MTZ. n = 17, K50 n = 7, 726 n = 19, 726+MTZ. n = 12. c Representative tumors post-harvest. d Representative image of lung metastases. Scale bar, 500 µm in the large image, 250 µm in the indicated inset. e Lung metastasis rank [number of metastases per lung area (pixel 2 )] calculated as percentage compared with the average metastasis rank in the vehicle control group of each experiment (set as 100% metastasis rank). Each symbol represents one mouse. Presented results are of three independent experiments. **** p = 4.08 × 10 −5 , * p = 0.0177, n.s.: non-significant, Holm’s correction, one-tailed Mann–Whitney. Vehicle n = 20, MTZ. n = 17, K50 n = 7, 726 n = 18, 726+met n = 12. f Images taken by fluorescence binocular of lung metastases in four mice implanted with GFP-expressing AT3 cells as described above. Two tumor-bearing mice were inoculated with F. nucleatum and two were treated by PBS vehicle. Scale bar, 1000 µm. Source data are provided as a Source data file.

    Article Snippet: Intravascularly inoculated Fap2-expressing F. nucleatum ATCC 23726 specifically colonize mice mammary tumors, whereas Fap2-deficient bacteria are impaired in tumor colonization.

    Techniques: Injection, Mouse Assay, One-tailed Test, MANN-WHITNEY, Fluorescence, Expressing

    F. nucleatum colonizes mammary cancer in a Fap2-mediated mechanism. a Orthotropic 4T1 BALB/c mouse mammary cancer model. When reaching 500 mm 3 size, 5 × 10 7 F. nucleatum ATCC 23726 were IV injected. Twenty-four hours post inoculation, breast cancer and normal tissue from an adjacent mammary were harvested. b Representative images and sum of fluorescence intensity of binding of FITC-labeled PNA (green) to tumor and normal tissue stained with Hoechst dye (blue). Each paired symbols represents one mouse ( n = 7), ** p = 0.0078, one-tailed, Wilcoxon matched-paired signed rank test. c Relative F. nucleatum gDNA abundance (2 −ΔCt ) in breast tissue from tumor-free mice (No tumor, n = 3), and in tumor and matching normal tissue of 4T1 tumor-bearing mice ( n = 5) all inoculated with F. nucleatum ATCC 23726. * p = 0.0358 Holm-corrected one-tailed Mann–Whitney test. * p = 0.0313, one-tailed, Wilcoxon matched-paired signed rank test (for paired normal adjacent-tumor samples). d Orthotropic AT3 C57BL/6 mouse mammary cancer model. When reaching 500 mm 3 , mice were inoculated with 5 × 10 7 F. nucleatum ATCC 23726 (726; blue), 5 × 10 7 Fap2 mutant K50 (MUT K50; red) or with 5 × 10 7 P. gingivalis (Pg; black). Twenty-four hours later, normal mammary and breast cancer tissue were harvested from each mice and bacteria quantified. e Bacterial abundance (CFU/g tissue), and relative bacterial gDNA abundance (2 −ΔCt ) in tumor and normal tissue of AT3 tumor-bearing mice inoculated with P. gingivalis ( Pg , n = 9), K50 ( n = 12), or F. nucleatum ATCC 23726 (726, n = 12). Each symbol represents one mouse. * p = 0.0156 one-tailed, Wilcoxon matched-paired signed rank test. *** p = 0.0006, ** p = 0.001598. Holm-corrected one-tailed Mann–Whitney test (left panel). **** p

    Journal: Nature Communications

    Article Title: Breast cancer colonization by Fusobacterium nucleatum accelerates tumor growth and metastatic progression

    doi: 10.1038/s41467-020-16967-2

    Figure Lengend Snippet: F. nucleatum colonizes mammary cancer in a Fap2-mediated mechanism. a Orthotropic 4T1 BALB/c mouse mammary cancer model. When reaching 500 mm 3 size, 5 × 10 7 F. nucleatum ATCC 23726 were IV injected. Twenty-four hours post inoculation, breast cancer and normal tissue from an adjacent mammary were harvested. b Representative images and sum of fluorescence intensity of binding of FITC-labeled PNA (green) to tumor and normal tissue stained with Hoechst dye (blue). Each paired symbols represents one mouse ( n = 7), ** p = 0.0078, one-tailed, Wilcoxon matched-paired signed rank test. c Relative F. nucleatum gDNA abundance (2 −ΔCt ) in breast tissue from tumor-free mice (No tumor, n = 3), and in tumor and matching normal tissue of 4T1 tumor-bearing mice ( n = 5) all inoculated with F. nucleatum ATCC 23726. * p = 0.0358 Holm-corrected one-tailed Mann–Whitney test. * p = 0.0313, one-tailed, Wilcoxon matched-paired signed rank test (for paired normal adjacent-tumor samples). d Orthotropic AT3 C57BL/6 mouse mammary cancer model. When reaching 500 mm 3 , mice were inoculated with 5 × 10 7 F. nucleatum ATCC 23726 (726; blue), 5 × 10 7 Fap2 mutant K50 (MUT K50; red) or with 5 × 10 7 P. gingivalis (Pg; black). Twenty-four hours later, normal mammary and breast cancer tissue were harvested from each mice and bacteria quantified. e Bacterial abundance (CFU/g tissue), and relative bacterial gDNA abundance (2 −ΔCt ) in tumor and normal tissue of AT3 tumor-bearing mice inoculated with P. gingivalis ( Pg , n = 9), K50 ( n = 12), or F. nucleatum ATCC 23726 (726, n = 12). Each symbol represents one mouse. * p = 0.0156 one-tailed, Wilcoxon matched-paired signed rank test. *** p = 0.0006, ** p = 0.001598. Holm-corrected one-tailed Mann–Whitney test (left panel). **** p

    Article Snippet: Intravascularly inoculated Fap2-expressing F. nucleatum ATCC 23726 specifically colonize mice mammary tumors, whereas Fap2-deficient bacteria are impaired in tumor colonization.

    Techniques: Injection, Fluorescence, Binding Assay, Labeling, Staining, One-tailed Test, Mouse Assay, MANN-WHITNEY, Mutagenesis

    Attachment of F. nucleatum ATCC 23726 to breast cancer is Fap2-mediated and Gal-GalNAc dependent. a Representative images (left panels, scale bar 20 µm) and quantitative analysis (Fn/mm 2 ) (right panel) of binding of Cy3-labeled (white) WT F. nucleatum ATCC 23726 (726) and of its Cy5-labeled (red) Fap2-inactivated isogenic mutant K50, to Hoechst-stained (blue) human breast cancer TMAs (HBre-Duc060CS-01 and BR1006). Results in the right panel are classified by tissue type (normal n = 35, benign n = 7, malignant n = 64) and bacterial strain (K50, 726). Each core is measured twice, once for each of the strains in the appropriate tissue type (as demonstrated in the pictures). When an individual contributed a single core to the experiment, the two observations are shown as circles (●); when she contributed two cores, these were to the normal type and the malignant type, and the four observations are depicted as plus signs (+). The six classes were compared against each other in pairs: the exact two-tailed Wilcoxon test was used for pairs that were from the same core, the exact two-tailed Mann–Whitney test for independent samples. Where the same hypothesis was tested on more than one pair, and the individual pairs were mutually independent, the corresponding p -values were combined by Fisher’s method. Wherever the pairs were not independent, the multiple comparisons were adjusted for using the Holm modification of the Bonferroni correction. **** p

    Journal: Nature Communications

    Article Title: Breast cancer colonization by Fusobacterium nucleatum accelerates tumor growth and metastatic progression

    doi: 10.1038/s41467-020-16967-2

    Figure Lengend Snippet: Attachment of F. nucleatum ATCC 23726 to breast cancer is Fap2-mediated and Gal-GalNAc dependent. a Representative images (left panels, scale bar 20 µm) and quantitative analysis (Fn/mm 2 ) (right panel) of binding of Cy3-labeled (white) WT F. nucleatum ATCC 23726 (726) and of its Cy5-labeled (red) Fap2-inactivated isogenic mutant K50, to Hoechst-stained (blue) human breast cancer TMAs (HBre-Duc060CS-01 and BR1006). Results in the right panel are classified by tissue type (normal n = 35, benign n = 7, malignant n = 64) and bacterial strain (K50, 726). Each core is measured twice, once for each of the strains in the appropriate tissue type (as demonstrated in the pictures). When an individual contributed a single core to the experiment, the two observations are shown as circles (●); when she contributed two cores, these were to the normal type and the malignant type, and the four observations are depicted as plus signs (+). The six classes were compared against each other in pairs: the exact two-tailed Wilcoxon test was used for pairs that were from the same core, the exact two-tailed Mann–Whitney test for independent samples. Where the same hypothesis was tested on more than one pair, and the individual pairs were mutually independent, the corresponding p -values were combined by Fisher’s method. Wherever the pairs were not independent, the multiple comparisons were adjusted for using the Holm modification of the Bonferroni correction. **** p

    Article Snippet: Intravascularly inoculated Fap2-expressing F. nucleatum ATCC 23726 specifically colonize mice mammary tumors, whereas Fap2-deficient bacteria are impaired in tumor colonization.

    Techniques: Binding Assay, Labeling, Mutagenesis, Staining, Two Tailed Test, MANN-WHITNEY, Modification

    Amino acid sequence alignment of FadA and its paralogues. Highlighted in gray are the identical residues shared among FadA proteins. The sequences of two FadA paralogues, FN1529 from F. nucleatum ATCC 25586 and FNV2159 from F. nucleatum ATCC 49256, are

    Journal: Journal of Bacteriology

    Article Title: Identification and Characterization of a Novel Adhesin Unique to Oral Fusobacteria

    doi: 10.1128/JB.187.15.5330-5340.2005

    Figure Lengend Snippet: Amino acid sequence alignment of FadA and its paralogues. Highlighted in gray are the identical residues shared among FadA proteins. The sequences of two FadA paralogues, FN1529 from F. nucleatum ATCC 25586 and FNV2159 from F. nucleatum ATCC 49256, are

    Article Snippet: The accession numbers for the FadA sequences from other fusobacterial strains and species are as follows: for F. nucleatum ATCC 10953 , for F. nucleatum ATCC 23726 , for F. nucleatum ATCC 25586 , for F. nucleatum ATCC 49256 , for F. nucleatum ATCC 51190 , for F. nucleatum DUMC1356 , for F. nucleatum DUMC2079 , for F. nucleatum DUMC2929 , for F. nucleatum DUMC3156 , for F. nucleatum DUMC3349 , for F. nucleatum PK1594 , for F. periodonticum ATCC 33693 , and for F. simiae ATCC 33568 .

    Techniques: Sequencing

    AT3 mammary tumor acceleration by F. nucleatum is immune-mediated. a 1 × 10 6 AT3 cells were injected to the mammary fat pad of 6–7-week-old female C57BL/6 mice. When tumor volume reached 500 mm 3 , mice were IV injected with PBS vehicle or with 5 × 10 7 F. nucleatum ATCC 23726 (726). At day 8 post injection, mice were sacrificed, tumors were harvested subjected to flow cytometry and abundance of NK cells, CD4+ T cells and CD8+ T cells was determined. Boxplot whiskers represent extrema, box bounds represent upper and lower quartiles, and center-line represents the median value. n.s.: non-significant. ** p = 0.00216, * p = 0.0152, two-tailed Mann–Whitney (each symbol represents the mean of two technical replicates, n = 6 per group). b 1 × 10 5 AT3 or GFP-expressing AT3 cells were injected to the mammary fat pad of 6–7-week-old female SCID-beige mice. When tumor volume reached 500 mm 3 , mice were IV injected with PBS vehicle or with 5 × 10 7 F. nucleatum ATCC 23726 (726). At day 8 post injection, mice were sacrificed, tumors were harvested and weighed in an analytic weigh. Boxplot whiskers represent extrema, box bounds represent upper and lower quartiles, and center-line represents the median value of two independent experiments. n.s.: non-significant. n = 8 per vehicle group and n = 10 per F. nucleatum -infected group. c Representative picture and densitometry analysis of gelatin zymography of conditioned medium prepared from mouse mammary tumor cell line AT3 or from the human breast cancer cell line MDA-MB-231 that were co-cultured or not, with F. nucleatum strains ATCC 25586 or ATCC 23726. Bars indicate mean ± SEM of six independent experiments. * p = 0.031 two-tailed paired Wilcoxon. Note that mouse MMP-9 is larger than that of human. Source data are provided as a Source data file.

    Journal: Nature Communications

    Article Title: Breast cancer colonization by Fusobacterium nucleatum accelerates tumor growth and metastatic progression

    doi: 10.1038/s41467-020-16967-2

    Figure Lengend Snippet: AT3 mammary tumor acceleration by F. nucleatum is immune-mediated. a 1 × 10 6 AT3 cells were injected to the mammary fat pad of 6–7-week-old female C57BL/6 mice. When tumor volume reached 500 mm 3 , mice were IV injected with PBS vehicle or with 5 × 10 7 F. nucleatum ATCC 23726 (726). At day 8 post injection, mice were sacrificed, tumors were harvested subjected to flow cytometry and abundance of NK cells, CD4+ T cells and CD8+ T cells was determined. Boxplot whiskers represent extrema, box bounds represent upper and lower quartiles, and center-line represents the median value. n.s.: non-significant. ** p = 0.00216, * p = 0.0152, two-tailed Mann–Whitney (each symbol represents the mean of two technical replicates, n = 6 per group). b 1 × 10 5 AT3 or GFP-expressing AT3 cells were injected to the mammary fat pad of 6–7-week-old female SCID-beige mice. When tumor volume reached 500 mm 3 , mice were IV injected with PBS vehicle or with 5 × 10 7 F. nucleatum ATCC 23726 (726). At day 8 post injection, mice were sacrificed, tumors were harvested and weighed in an analytic weigh. Boxplot whiskers represent extrema, box bounds represent upper and lower quartiles, and center-line represents the median value of two independent experiments. n.s.: non-significant. n = 8 per vehicle group and n = 10 per F. nucleatum -infected group. c Representative picture and densitometry analysis of gelatin zymography of conditioned medium prepared from mouse mammary tumor cell line AT3 or from the human breast cancer cell line MDA-MB-231 that were co-cultured or not, with F. nucleatum strains ATCC 25586 or ATCC 23726. Bars indicate mean ± SEM of six independent experiments. * p = 0.031 two-tailed paired Wilcoxon. Note that mouse MMP-9 is larger than that of human. Source data are provided as a Source data file.

    Article Snippet: Bacterial strains and growth conditions F. nucleatum strains ATCC 25586, ATCC 23726, the isogenic Fap2-inactivated mutants K50, D22 and P. gingivalis ATCC 33277 were grown in Wilkins Chalgren broth (Oxoid, UK) or on Columbia agar plates (Oxoid, UK) supplemented with 5% defibrinated sheep blood (Novamed, Israel).

    Techniques: Injection, Mouse Assay, Flow Cytometry, Two Tailed Test, MANN-WHITNEY, Expressing, Infection, Zymography, Multiple Displacement Amplification, Cell Culture

    A model of F. nucleatum induced metastasis through cytokine signaling. We show that F. nucleatum induces IL-8 and CXCL1 secretion from CRC cells, and both cytokines have been characterized as key players in cancer metastasis and subsequent downstream cell seeding. As well as autocrine signaling back to cancer cells as a metastatic signal, HCT116 derived cytokines can participate in paracrine signaling to recruit neighboring immune cells, which further secrete their own cytokine signatures that alter the tumor microenvironment through metastatic, inflammatory, and immune cell programming.

    Journal: bioRxiv

    Article Title: Fusobacterium nucleatum host cell binding and invasion induces IL-8 and CXCL1 secretion that drives colorectal cancer cell migration

    doi: 10.1101/2020.01.15.907931

    Figure Lengend Snippet: A model of F. nucleatum induced metastasis through cytokine signaling. We show that F. nucleatum induces IL-8 and CXCL1 secretion from CRC cells, and both cytokines have been characterized as key players in cancer metastasis and subsequent downstream cell seeding. As well as autocrine signaling back to cancer cells as a metastatic signal, HCT116 derived cytokines can participate in paracrine signaling to recruit neighboring immune cells, which further secrete their own cytokine signatures that alter the tumor microenvironment through metastatic, inflammatory, and immune cell programming.

    Article Snippet: Cells were loaded onto a Guava easyCyte 5 flow cytometer (Luminex) and 10,000 cells were collected using an initial single cell gate to measure the median green fluorescence induced by intracellular F. nucleatum labeled with FM 1-43FX.

    Techniques: Derivative Assay

    Validation of markerless fap2 and fadA gene deletions in F. nucleatum 23726 Δ galKT. ( A ) Map of chromosomal plasmid incorporation after the initial single-crossover insertion for fap2 . ( B ) RT-PCR of fadA and fap2 with their two surrounding genes showing loss of gene transcription due to gene deletion. ( C ) DNA sequencing with primers sitting −250 and +250 from the start codon of fadA and fap2 . All constructs created are 100% accurate and show perfect excision and recission of the genome to the −1 and +1 base for each gene.

    Journal: bioRxiv

    Article Title: Fusobacterium nucleatum host cell binding and invasion induces IL-8 and CXCL1 secretion that drives colorectal cancer cell migration

    doi: 10.1101/2020.01.15.907931

    Figure Lengend Snippet: Validation of markerless fap2 and fadA gene deletions in F. nucleatum 23726 Δ galKT. ( A ) Map of chromosomal plasmid incorporation after the initial single-crossover insertion for fap2 . ( B ) RT-PCR of fadA and fap2 with their two surrounding genes showing loss of gene transcription due to gene deletion. ( C ) DNA sequencing with primers sitting −250 and +250 from the start codon of fadA and fap2 . All constructs created are 100% accurate and show perfect excision and recission of the genome to the −1 and +1 base for each gene.

    Article Snippet: Cells were loaded onto a Guava easyCyte 5 flow cytometer (Luminex) and 10,000 cells were collected using an initial single cell gate to measure the median green fluorescence induced by intracellular F. nucleatum labeled with FM 1-43FX.

    Techniques: Plasmid Preparation, Reverse Transcription Polymerase Chain Reaction, DNA Sequencing, Construct

    Utilizing the Leloir pathway for targeted gene deletions in bacteria. ( A ) The Leloir pathway of galactose catabolism. ( B ) Graphical representation of the different selection stages of F. nucleatum gene deletions and how the GalK and GalT affect cellular survival in the presence of galactose and 2-deoxy-galactose.

    Journal: bioRxiv

    Article Title: Fusobacterium nucleatum host cell binding and invasion induces IL-8 and CXCL1 secretion that drives colorectal cancer cell migration

    doi: 10.1101/2020.01.15.907931

    Figure Lengend Snippet: Utilizing the Leloir pathway for targeted gene deletions in bacteria. ( A ) The Leloir pathway of galactose catabolism. ( B ) Graphical representation of the different selection stages of F. nucleatum gene deletions and how the GalK and GalT affect cellular survival in the presence of galactose and 2-deoxy-galactose.

    Article Snippet: Cells were loaded onto a Guava easyCyte 5 flow cytometer (Luminex) and 10,000 cells were collected using an initial single cell gate to measure the median green fluorescence induced by intracellular F. nucleatum labeled with FM 1-43FX.

    Techniques: Selection

    Deletion of the galKT gene operon in F. nucleatum 23726. ( A ) Development of pDJSVT13, the plasmid for galKT gene deletion, by inserting 1000 bp galKT flanking sequences into pDJSVT1. ( B ) Representation of single-crossover homologous recombination onto the F. nucleatum 23726 chromosome with either the A (upstream) or ( C ) B (downstream) 1000 bp DNA fragments in pDJSVT13 followed by showing how the double crossover results in complete plasmid excision. ( D ) Map of chromosomal plasmid incorporation after the initial single-crossover insertion. ( E ) PCR verification of single-cross pDJSVT13 and subsequent double-crossover deletion of the galKT operon. ( F ) Plating on 5 μg/mL thiamphenicol (T5) (single-crossover selection) and ( G ) 1% 2-deoxy-galactose (double-crossover), resulting in galKT operon deletion. ( H ) Validation that F. nucleatum 23726 Δ galKT grows the same as WT F. nucleatum 23726.

    Journal: bioRxiv

    Article Title: Fusobacterium nucleatum host cell binding and invasion induces IL-8 and CXCL1 secretion that drives colorectal cancer cell migration

    doi: 10.1101/2020.01.15.907931

    Figure Lengend Snippet: Deletion of the galKT gene operon in F. nucleatum 23726. ( A ) Development of pDJSVT13, the plasmid for galKT gene deletion, by inserting 1000 bp galKT flanking sequences into pDJSVT1. ( B ) Representation of single-crossover homologous recombination onto the F. nucleatum 23726 chromosome with either the A (upstream) or ( C ) B (downstream) 1000 bp DNA fragments in pDJSVT13 followed by showing how the double crossover results in complete plasmid excision. ( D ) Map of chromosomal plasmid incorporation after the initial single-crossover insertion. ( E ) PCR verification of single-cross pDJSVT13 and subsequent double-crossover deletion of the galKT operon. ( F ) Plating on 5 μg/mL thiamphenicol (T5) (single-crossover selection) and ( G ) 1% 2-deoxy-galactose (double-crossover), resulting in galKT operon deletion. ( H ) Validation that F. nucleatum 23726 Δ galKT grows the same as WT F. nucleatum 23726.

    Article Snippet: Cells were loaded onto a Guava easyCyte 5 flow cytometer (Luminex) and 10,000 cells were collected using an initial single cell gate to measure the median green fluorescence induced by intracellular F. nucleatum labeled with FM 1-43FX.

    Techniques: Plasmid Preparation, Homologous Recombination, Polymerase Chain Reaction, Selection

    Development of vector for markerless gene deletion in Fusobacterium nucleatum . ( A ) Method to create pDJSVT1; the base vector for all gene deletion constructs that consists of an E. coli origin and chloramphenicol/thiamphenicol resistance and a GC rich multiple cloning site for efficient cloning of AT rich F. nucleatum DNA. ( B ) pDJSVT7 plasmid incorporation of a constitutively active FLAG:galK gene for selection on 2-deoxy-galactose. ( C ) Development of a chromosomal complementation vector that incorporates the entire pDJSVT11 plasmid onto the chromosome at the arsB gene in F. nucleatum 23726.

    Journal: bioRxiv

    Article Title: Fusobacterium nucleatum host cell binding and invasion induces IL-8 and CXCL1 secretion that drives colorectal cancer cell migration

    doi: 10.1101/2020.01.15.907931

    Figure Lengend Snippet: Development of vector for markerless gene deletion in Fusobacterium nucleatum . ( A ) Method to create pDJSVT1; the base vector for all gene deletion constructs that consists of an E. coli origin and chloramphenicol/thiamphenicol resistance and a GC rich multiple cloning site for efficient cloning of AT rich F. nucleatum DNA. ( B ) pDJSVT7 plasmid incorporation of a constitutively active FLAG:galK gene for selection on 2-deoxy-galactose. ( C ) Development of a chromosomal complementation vector that incorporates the entire pDJSVT11 plasmid onto the chromosome at the arsB gene in F. nucleatum 23726.

    Article Snippet: Cells were loaded onto a Guava easyCyte 5 flow cytometer (Luminex) and 10,000 cells were collected using an initial single cell gate to measure the median green fluorescence induced by intracellular F. nucleatum labeled with FM 1-43FX.

    Techniques: Plasmid Preparation, Construct, Clone Assay, Selection

    F. nucleatum induces cytokine secretion from HCT116 cells. ( A ) Overview of experiments used to analyze Fnn induced cytokine secretion from HCT116 CRC cells. ( B ) Broad cytokine array dot blots analyzing Fnn and Fnn adhesin deletion strains. ( C ) Heatmap of fold increased dot blot intensity reveals an increase in IL-8 and CXCL1 secretion for invasive strains. ( D ) IL-8 and CXCL1 ELISA to quantitate cytokine secretion from HCT116 cells induced by Fnn and Fnn Δ fap2. ( E ) IL-8 and CXCL1 ELISA to quantitate and compare cytokine secretion from HCT116 cells induced by F. nucleatum subsp. nucleatum 25586, F. nucleatum subsp. nucleatum 23726, F. nucleatum subsp. animalis 7_1 ( Fna ).

    Journal: bioRxiv

    Article Title: Fusobacterium nucleatum host cell binding and invasion induces IL-8 and CXCL1 secretion that drives colorectal cancer cell migration

    doi: 10.1101/2020.01.15.907931

    Figure Lengend Snippet: F. nucleatum induces cytokine secretion from HCT116 cells. ( A ) Overview of experiments used to analyze Fnn induced cytokine secretion from HCT116 CRC cells. ( B ) Broad cytokine array dot blots analyzing Fnn and Fnn adhesin deletion strains. ( C ) Heatmap of fold increased dot blot intensity reveals an increase in IL-8 and CXCL1 secretion for invasive strains. ( D ) IL-8 and CXCL1 ELISA to quantitate cytokine secretion from HCT116 cells induced by Fnn and Fnn Δ fap2. ( E ) IL-8 and CXCL1 ELISA to quantitate and compare cytokine secretion from HCT116 cells induced by F. nucleatum subsp. nucleatum 25586, F. nucleatum subsp. nucleatum 23726, F. nucleatum subsp. animalis 7_1 ( Fna ).

    Article Snippet: Cells were loaded onto a Guava easyCyte 5 flow cytometer (Luminex) and 10,000 cells were collected using an initial single cell gate to measure the median green fluorescence induced by intracellular F. nucleatum labeled with FM 1-43FX.

    Techniques: Dot Blot, Enzyme-linked Immunosorbent Assay

    Target gene deletion in F. nucleatum 23726 Δ galKT. ( A ) Development of all target gene deletion plasmids ( Table S2 ) by inserting 750 bp from target gene flanking sequences into pDJSVT7. ( B ) Representation of single-crossover homologous recombination onto the F. nucleatum 23726 chromosome with either the A (upstream) or ( C ) B (downstream) 750 bp DNA fragments. Double crossover results are shown during complete plasmid excision. ( D ) fap2 and fadA as examples of gene excision. KO validation primers are shown flanking genes at −1000 and +1000 bp. This results in 2kb PCR products as shown in ( E ) for the fap2 and fadA gene deletions. ( F ) Table of select deleted genes in this study and their function.

    Journal: bioRxiv

    Article Title: Fusobacterium nucleatum host cell binding and invasion induces IL-8 and CXCL1 secretion that drives colorectal cancer cell migration

    doi: 10.1101/2020.01.15.907931

    Figure Lengend Snippet: Target gene deletion in F. nucleatum 23726 Δ galKT. ( A ) Development of all target gene deletion plasmids ( Table S2 ) by inserting 750 bp from target gene flanking sequences into pDJSVT7. ( B ) Representation of single-crossover homologous recombination onto the F. nucleatum 23726 chromosome with either the A (upstream) or ( C ) B (downstream) 750 bp DNA fragments. Double crossover results are shown during complete plasmid excision. ( D ) fap2 and fadA as examples of gene excision. KO validation primers are shown flanking genes at −1000 and +1000 bp. This results in 2kb PCR products as shown in ( E ) for the fap2 and fadA gene deletions. ( F ) Table of select deleted genes in this study and their function.

    Article Snippet: Cells were loaded onto a Guava easyCyte 5 flow cytometer (Luminex) and 10,000 cells were collected using an initial single cell gate to measure the median green fluorescence induced by intracellular F. nucleatum labeled with FM 1-43FX.

    Techniques: Homologous Recombination, Plasmid Preparation, Polymerase Chain Reaction

    F. nucleatum binds to and invades HCT116 cells. ( A ) Overview of experiments used to analyze binding and invasion of Fnn and adhesin mutants in HCT116 CRC cells. ( B ) Intra- and extracellular Fnn detected with a fluorescent lipid dye (Green, intra- and extracellular) and a pan- Fusobacterium membrane antisera (Red, extracellular), host cell nuclei are stained with DAPI (blue). ( C ) Zoomed in view and focus on a single Fnn bacterium that is half intracellular, half extracellular. ( D ) Imaging flow cytometry view of intracellular Fnn . ( E ) Comparison of HCT116 binding and invasion of wild type (WT) F. nucleatum 23726 with F. nucleatum 23726 Δ galKT ( Fnn ). ( F ) Binding and invasion analysis of Fnn and adhesin gene deletion strains using flow cytometry. ( G ) Invasion and survival of Fnn and Fnn Δ fap2 in HCT116 cells analyzed using an antibiotic protection assay.

    Journal: bioRxiv

    Article Title: Fusobacterium nucleatum host cell binding and invasion induces IL-8 and CXCL1 secretion that drives colorectal cancer cell migration

    doi: 10.1101/2020.01.15.907931

    Figure Lengend Snippet: F. nucleatum binds to and invades HCT116 cells. ( A ) Overview of experiments used to analyze binding and invasion of Fnn and adhesin mutants in HCT116 CRC cells. ( B ) Intra- and extracellular Fnn detected with a fluorescent lipid dye (Green, intra- and extracellular) and a pan- Fusobacterium membrane antisera (Red, extracellular), host cell nuclei are stained with DAPI (blue). ( C ) Zoomed in view and focus on a single Fnn bacterium that is half intracellular, half extracellular. ( D ) Imaging flow cytometry view of intracellular Fnn . ( E ) Comparison of HCT116 binding and invasion of wild type (WT) F. nucleatum 23726 with F. nucleatum 23726 Δ galKT ( Fnn ). ( F ) Binding and invasion analysis of Fnn and adhesin gene deletion strains using flow cytometry. ( G ) Invasion and survival of Fnn and Fnn Δ fap2 in HCT116 cells analyzed using an antibiotic protection assay.

    Article Snippet: Cells were loaded onto a Guava easyCyte 5 flow cytometer (Luminex) and 10,000 cells were collected using an initial single cell gate to measure the median green fluorescence induced by intracellular F. nucleatum labeled with FM 1-43FX.

    Techniques: Binding Assay, Staining, Imaging, Flow Cytometry

    Complementation of gene deletions at the static arsB gene site in F. nucleatum 23726. ( A ) pDJSVT20 plasmid created for complementing Δ galKT with galKT::6xHis. ( B ) RT-PCR of Δ galKT and Δ galKT galKT::6xHis and the two genes flanking the operon (FN2106, FN2109). ( C ) Western blot verification of galKT::6sHis complementation and constitutive expression of GalT::6xHis. ( D ) pDJSVT11 base vector depicted with target gene of interest (GOI) complementation and how it incorporates into the chromosome at arsB . ( E ) Potential sites for complementation of gene deletions used in this study. However, we did not complement the Δ fap2 strain due to the extreme difficulties of complementing large genes (12kb). ( F ) Complementation of Δ fadA with fadA: : FLAG and western blot verification of constitutive expression of FadA::FLAG.

    Journal: bioRxiv

    Article Title: Fusobacterium nucleatum host cell binding and invasion induces IL-8 and CXCL1 secretion that drives colorectal cancer cell migration

    doi: 10.1101/2020.01.15.907931

    Figure Lengend Snippet: Complementation of gene deletions at the static arsB gene site in F. nucleatum 23726. ( A ) pDJSVT20 plasmid created for complementing Δ galKT with galKT::6xHis. ( B ) RT-PCR of Δ galKT and Δ galKT galKT::6xHis and the two genes flanking the operon (FN2106, FN2109). ( C ) Western blot verification of galKT::6sHis complementation and constitutive expression of GalT::6xHis. ( D ) pDJSVT11 base vector depicted with target gene of interest (GOI) complementation and how it incorporates into the chromosome at arsB . ( E ) Potential sites for complementation of gene deletions used in this study. However, we did not complement the Δ fap2 strain due to the extreme difficulties of complementing large genes (12kb). ( F ) Complementation of Δ fadA with fadA: : FLAG and western blot verification of constitutive expression of FadA::FLAG.

    Article Snippet: Cells were loaded onto a Guava easyCyte 5 flow cytometer (Luminex) and 10,000 cells were collected using an initial single cell gate to measure the median green fluorescence induced by intracellular F. nucleatum labeled with FM 1-43FX.

    Techniques: Plasmid Preparation, Reverse Transcription Polymerase Chain Reaction, Western Blot, Expressing