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  • 99
    ATCC f nucleatum atcc 25586
    F Nucleatum Atcc 25586, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 692 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Scientific Culti Loops are ready to use QC organisms recommended for use in performance testing of media stains reagents and identification kits and for the evaluation of bacteriological procedures
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    99
    ATCC f nucleatum atcc 23726
    Roles of fusobacterial ftsX and envC in the development of monospecies and multispecies biofilms. (A) ftsX locus in the chromosome of F. <t>nucleatum</t> ATCC 23726, with envC adjacent to ppnK , which encodes an NAD+ kinase. Upstream of ftsX is fadA ); expression of genes in the ftsX locus and fadA appears to be controlled by individual promoters. (B) Map of the nonreplicative vector pCWU8 used to generate unmarked, in-frame gene deletion mutants in F. nucleatum . The kanamycin resistance cassette ( kan ), chloramphenicol/thiamphenicol resistance gene ( catP ), and galK are indicated. The multiple cloning sites (MCS) contain EcoRI, SacI, KpnI, BamHI, and SalI. (C) Fusobacterial strains were evaluated for their ability to form monospecies biofilm using crystal violet staining. (D) Interaction of fusobacteria and S. oralis So34 was determined by a standard coaggregation assay, with a radD mutant used as a negative control. (E) Three-species biofilms were analyzed by confocal laser scanning microscopy at a magnification of ×20, using 16S rRNA-oligonucleotide probes specific for F. nucleatum (red), A. oris (green), and S. oralis (blue); side and top views are presented. The results are representative of three independent experiments performed in triplicate.
    F Nucleatum Atcc 23726, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 385 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    ATCC f nucleatum atcc 10953
    Restriction maps and Southern blots of the F. <t>nucleatum</t> plasmids. (A) Partial restriction map of F. nucleatum plasmids pFN1, pFN2 and pFN3 and shuttle plasmid pHS17. Selected restriction endonuclease sites in the native plasmids are presented. Restriction endonuclease sites indicated for pHS17 relate to the plasmid construction. The pFN1 portion of pHS17 is indicated by the thick solid bar, with the position of the repA homologue (ORF5) and putative ori indicated. (B and C) Plasmids from F. nucleatum strains 12230 (pFN1, lanes 1), 10113 (pFN2, lanes 2), and <t>ATCC</t> 10953 (pFN3, lanes 3) were probed with pFN1 DNA with Eco RI digests (B) or pFN3 DNA with Eco RV digests (C). The Hin cII-digested pFN1 and Ase I-digested pFN3 probes were 32 P labeled (specific activity of 25 × 10 8 and 4.5 × 10 8 dpm/μg of DNA, respectively). The positions of molecular size markers are indicated on the left, and the linear forms of pFN1, pFN2, and pFN3 are indicated on the right.
    F Nucleatum Atcc 10953, supplied by ATCC, used in various techniques. Bioz Stars score: 95/100, based on 244 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    ATCC f nucleatum atcc 49256
    Restriction maps and Southern blots of the F. <t>nucleatum</t> plasmids. (A) Partial restriction map of F. nucleatum plasmids pFN1, pFN2 and pFN3 and shuttle plasmid pHS17. Selected restriction endonuclease sites in the native plasmids are presented. Restriction endonuclease sites indicated for pHS17 relate to the plasmid construction. The pFN1 portion of pHS17 is indicated by the thick solid bar, with the position of the repA homologue (ORF5) and putative ori indicated. (B and C) Plasmids from F. nucleatum strains 12230 (pFN1, lanes 1), 10113 (pFN2, lanes 2), and <t>ATCC</t> 10953 (pFN3, lanes 3) were probed with pFN1 DNA with Eco RI digests (B) or pFN3 DNA with Eco RV digests (C). The Hin cII-digested pFN1 and Ase I-digested pFN3 probes were 32 P labeled (specific activity of 25 × 10 8 and 4.5 × 10 8 dpm/μg of DNA, respectively). The positions of molecular size markers are indicated on the left, and the linear forms of pFN1, pFN2, and pFN3 are indicated on the right.
    F Nucleatum Atcc 49256, supplied by ATCC, used in various techniques. Bioz Stars score: 94/100, based on 75 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    ATCC f nucleatum strains atcc 25586
    AT3 mammary tumor acceleration by F. <t>nucleatum</t> 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.
    F Nucleatum Strains Atcc 25586, supplied by ATCC, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    ATCC × 107 f nucleatum atcc 23726
    AT3 mammary tumor acceleration by F. <t>nucleatum</t> 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.
    × 107 F Nucleatum Atcc 23726, supplied by ATCC, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    86
    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: 86/100, based on 13 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    ATCC f nucleatum atcc 25286
    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 25286, supplied by ATCC, used in various techniques. Bioz Stars score: 94/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    ATCC fitc labeled 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.
    Fitc Labeled F Nucleatum Atcc 23726, supplied by ATCC, used in various techniques. Bioz Stars score: 88/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    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
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    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
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    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
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    Image Search Results


    Roles of fusobacterial ftsX and envC in the development of monospecies and multispecies biofilms. (A) ftsX locus in the chromosome of F. nucleatum ATCC 23726, with envC adjacent to ppnK , which encodes an NAD+ kinase. Upstream of ftsX is fadA ); expression of genes in the ftsX locus and fadA appears to be controlled by individual promoters. (B) Map of the nonreplicative vector pCWU8 used to generate unmarked, in-frame gene deletion mutants in F. nucleatum . The kanamycin resistance cassette ( kan ), chloramphenicol/thiamphenicol resistance gene ( catP ), and galK are indicated. The multiple cloning sites (MCS) contain EcoRI, SacI, KpnI, BamHI, and SalI. (C) Fusobacterial strains were evaluated for their ability to form monospecies biofilm using crystal violet staining. (D) Interaction of fusobacteria and S. oralis So34 was determined by a standard coaggregation assay, with a radD mutant used as a negative control. (E) Three-species biofilms were analyzed by confocal laser scanning microscopy at a magnification of ×20, using 16S rRNA-oligonucleotide probes specific for F. nucleatum (red), A. oris (green), and S. oralis (blue); side and top views are presented. The results are representative of three independent experiments performed in triplicate.

    Journal: mBio

    Article Title: Forward Genetic Dissection of Biofilm Development by Fusobacterium nucleatum: Novel Functions of Cell Division Proteins FtsX and EnvC

    doi: 10.1128/mBio.00360-18

    Figure Lengend Snippet: Roles of fusobacterial ftsX and envC in the development of monospecies and multispecies biofilms. (A) ftsX locus in the chromosome of F. nucleatum ATCC 23726, with envC adjacent to ppnK , which encodes an NAD+ kinase. Upstream of ftsX is fadA ); expression of genes in the ftsX locus and fadA appears to be controlled by individual promoters. (B) Map of the nonreplicative vector pCWU8 used to generate unmarked, in-frame gene deletion mutants in F. nucleatum . The kanamycin resistance cassette ( kan ), chloramphenicol/thiamphenicol resistance gene ( catP ), and galK are indicated. The multiple cloning sites (MCS) contain EcoRI, SacI, KpnI, BamHI, and SalI. (C) Fusobacterial strains were evaluated for their ability to form monospecies biofilm using crystal violet staining. (D) Interaction of fusobacteria and S. oralis So34 was determined by a standard coaggregation assay, with a radD mutant used as a negative control. (E) Three-species biofilms were analyzed by confocal laser scanning microscopy at a magnification of ×20, using 16S rRNA-oligonucleotide probes specific for F. nucleatum (red), A. oris (green), and S. oralis (blue); side and top views are presented. The results are representative of three independent experiments performed in triplicate.

    Article Snippet: The DNA fragment compassing ftsX and envC was PCR amplified from the genomic DNA of F. nucleatum ATCC 23726 using primers ftsX-F2 and com-EnvC-R.

    Techniques: Expressing, Plasmid Preparation, Clone Assay, Staining, Mutagenesis, Negative Control, Confocal Laser Scanning Microscopy

    Roles of fusobacterial ftsX and envC in the development of monospecies and multispecies biofilms. (A) ftsX locus in the chromosome of F. nucleatum ATCC 23726, with envC adjacent to ppnK , which encodes an NAD+ kinase. Upstream of ftsX is fadA coding for the adhesin FadA ( 6 ); expression of genes in the ftsX locus and fadA appears to be controlled by individual promoters. (B) Map of the nonreplicative vector pCWU8 used to generate unmarked, in-frame gene deletion mutants in F. nucleatum . The kanamycin resistance cassette ( kan ), chloramphenicol/thiamphenicol resistance gene ( catP ), and galK are indicated. The multiple cloning sites (MCS) contain EcoRI, SacI, KpnI, BamHI, and SalI. (C) Fusobacterial strains were evaluated for their ability to form monospecies biofilm using crystal violet staining. (D) Interaction of fusobacteria and S. oralis So34 was determined by a standard coaggregation assay, with a radD mutant used as a negative control. (E) Three-species biofilms were analyzed by confocal laser scanning microscopy at a magnification of ×20, using 16S rRNA-oligonucleotide probes specific for F. nucleatum (red), A. oris (green), and S. oralis (blue); side and top views are presented. The results are representative of three independent experiments performed in triplicate.

    Journal: mBio

    Article Title: Forward Genetic Dissection of Biofilm Development by Fusobacterium nucleatum: Novel Functions of Cell Division Proteins FtsX and EnvC

    doi: 10.1128/mBio.00360-18

    Figure Lengend Snippet: Roles of fusobacterial ftsX and envC in the development of monospecies and multispecies biofilms. (A) ftsX locus in the chromosome of F. nucleatum ATCC 23726, with envC adjacent to ppnK , which encodes an NAD+ kinase. Upstream of ftsX is fadA coding for the adhesin FadA ( 6 ); expression of genes in the ftsX locus and fadA appears to be controlled by individual promoters. (B) Map of the nonreplicative vector pCWU8 used to generate unmarked, in-frame gene deletion mutants in F. nucleatum . The kanamycin resistance cassette ( kan ), chloramphenicol/thiamphenicol resistance gene ( catP ), and galK are indicated. The multiple cloning sites (MCS) contain EcoRI, SacI, KpnI, BamHI, and SalI. (C) Fusobacterial strains were evaluated for their ability to form monospecies biofilm using crystal violet staining. (D) Interaction of fusobacteria and S. oralis So34 was determined by a standard coaggregation assay, with a radD mutant used as a negative control. (E) Three-species biofilms were analyzed by confocal laser scanning microscopy at a magnification of ×20, using 16S rRNA-oligonucleotide probes specific for F. nucleatum (red), A. oris (green), and S. oralis (blue); side and top views are presented. The results are representative of three independent experiments performed in triplicate.

    Article Snippet: A library of approximately 24,000 thiamphenicol-resistant Tn5 mutants were generated from the parental F. nucleatum ATCC 23726 strain from a single Tn5 transposome reaction that was carried out in bulk, based on reaction efficiency determined from small-scale quantitative electroporation experiments.

    Techniques: Expressing, Plasmid Preparation, Clone Assay, Staining, Mutagenesis, Negative Control, Confocal Laser Scanning Microscopy

    Restriction maps and Southern blots of the F. nucleatum plasmids. (A) Partial restriction map of F. nucleatum plasmids pFN1, pFN2 and pFN3 and shuttle plasmid pHS17. Selected restriction endonuclease sites in the native plasmids are presented. Restriction endonuclease sites indicated for pHS17 relate to the plasmid construction. The pFN1 portion of pHS17 is indicated by the thick solid bar, with the position of the repA homologue (ORF5) and putative ori indicated. (B and C) Plasmids from F. nucleatum strains 12230 (pFN1, lanes 1), 10113 (pFN2, lanes 2), and ATCC 10953 (pFN3, lanes 3) were probed with pFN1 DNA with Eco RI digests (B) or pFN3 DNA with Eco RV digests (C). The Hin cII-digested pFN1 and Ase I-digested pFN3 probes were 32 P labeled (specific activity of 25 × 10 8 and 4.5 × 10 8 dpm/μg of DNA, respectively). The positions of molecular size markers are indicated on the left, and the linear forms of pFN1, pFN2, and pFN3 are indicated on the right.

    Journal: Journal of Bacteriology

    Article Title: Native Plasmids of Fusobacterium nucleatum: Characterization and Use in Development of Genetic Systems †

    doi:

    Figure Lengend Snippet: Restriction maps and Southern blots of the F. nucleatum plasmids. (A) Partial restriction map of F. nucleatum plasmids pFN1, pFN2 and pFN3 and shuttle plasmid pHS17. Selected restriction endonuclease sites in the native plasmids are presented. Restriction endonuclease sites indicated for pHS17 relate to the plasmid construction. The pFN1 portion of pHS17 is indicated by the thick solid bar, with the position of the repA homologue (ORF5) and putative ori indicated. (B and C) Plasmids from F. nucleatum strains 12230 (pFN1, lanes 1), 10113 (pFN2, lanes 2), and ATCC 10953 (pFN3, lanes 3) were probed with pFN1 DNA with Eco RI digests (B) or pFN3 DNA with Eco RV digests (C). The Hin cII-digested pFN1 and Ase I-digested pFN3 probes were 32 P labeled (specific activity of 25 × 10 8 and 4.5 × 10 8 dpm/μg of DNA, respectively). The positions of molecular size markers are indicated on the left, and the linear forms of pFN1, pFN2, and pFN3 are indicated on the right.

    Article Snippet: A shuttle plasmid, pHS17, capable of transforming Escherichia coli and F. nucleatum ATCC 10953 was constructed with pFN1. pHS17 was stably maintained in the F. nucleatum transformants, and differences in the transformation efficiencies suggested the presence of a restriction-modification system in F. nucleatum .

    Techniques: Plasmid Preparation, Labeling, Activity 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

    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: F. nucleatum strains ATCC 25586 and ATCC 23726 were brought to OD600 = 1 and added to the cell cultures at cell to bacterial ratio of 1:100.

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

    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