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  • 98
    Millipore sialidase activity
    Neu1 and Neu3 on sperm and change during capacitation. A , fluorescent staining of Neu1 on mouse sperm head before and after capacitation and counterstained with DAPI and an unspecific IgG control. The change was quantified using FACS and could be partially inhibited by DANA. B , fluorescent staining of Neu3 on mouse sperm head before and after capacitation and counterstained with DAPI and an unspecific IgG control. The change was quantified using FACS and could be partially inhibited by DANA. A and B are representative of 12 experiments each. C , Neu1/3 on mouse sperm in the testes and the proximal and distal epididymis. Arrowheads show positive staining only in mature sperm of the cauda (distal epididymis). HE , hematoxylin and eosin. D , NEU1/3 on human ejaculated sperm. Expression was highest in the hind part of the sperm head. The counterstain is DAPI ( blue ). ALF488 , Alexa Fluor 488. E , release of Neu1 and Neu3 from sperm into the supernatant and the effect of the <t>sialidase</t> inhibitor DANA. U , uncapacitated; C , capacitated; C + I , capacitated with the inhibitor DANA. F , sialidase activity in and release from sperm are both affected by protein inhibitor ( PI ). ***, p ≤ 0.001 versus the control.
    Sialidase Activity, supplied by Millipore, used in various techniques. Bioz Stars score: 98/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    86
    Thermo Fisher sialidase activity
    <t>Sialidase</t> activity of T. forsythia NanH and SiaHI expressed in E. coli . Strains KCL116 ( E. coli pET30; lanes 1 and 2), KCL117 [ E. coli BL21(DE3)/pET30:: nanH ; lanes 3 and 4], and KCL120 [ E. coli BL21(DE3)/pET30:: siaHI ; lanes 5 and 6] were grown in LB until
    Sialidase Activity, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sialidase activity/product/Thermo Fisher
    Average 86 stars, based on 1 article reviews
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    sialidase activity - by Bioz Stars, 2021-09
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    93
    Envigo sialidase activity
    Human vaginal microbial communities-collection and amplification. Step 1 - Anaerobic and aerobic vaginal swabs were collected on the same day from each participant. Aerobic swabs were eluted in sodium acetate buffer (pH 5.5) and <t>sialidase</t> activity was checked in the swab eluates using fluorogenic 4MU-Neu5Ac substrate. Anaerobic swabs were eluted in 2X NYCIII media (in an anaerobic chamber) and the communities were “fresh frozen”, without any amplification / overnight culture , by mixing with cryoprotectant and storing at −80°C. Step 2 - On the day of the experiment - fresh frozen anaerobic vaginal communities, from women who had detectable sialidase activity in their aerobic swab eluates, were thawed at 4°C and diluted 4-fold in NYCIII media (in an anaerobic chamber). The diluted communities were used for co-culture experiments with F. nucleatum .
    Sialidase Activity, supplied by Envigo, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    96
    Thermo Fisher na fluor influenza neuraminidase assay kit
    Human vaginal microbial communities-collection and amplification. Step 1 - Anaerobic and aerobic vaginal swabs were collected on the same day from each participant. Aerobic swabs were eluted in sodium acetate buffer (pH 5.5) and <t>sialidase</t> activity was checked in the swab eluates using fluorogenic 4MU-Neu5Ac substrate. Anaerobic swabs were eluted in 2X NYCIII media (in an anaerobic chamber) and the communities were “fresh frozen”, without any amplification / overnight culture , by mixing with cryoprotectant and storing at −80°C. Step 2 - On the day of the experiment - fresh frozen anaerobic vaginal communities, from women who had detectable sialidase activity in their aerobic swab eluates, were thawed at 4°C and diluted 4-fold in NYCIII media (in an anaerobic chamber). The diluted communities were used for co-culture experiments with F. nucleatum .
    Na Fluor Influenza Neuraminidase Assay Kit, supplied by Thermo Fisher, 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/na fluor influenza neuraminidase assay kit/product/Thermo Fisher
    Average 96 stars, based on 1 article reviews
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    na fluor influenza neuraminidase assay kit - by Bioz Stars, 2021-09
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    Image Search Results


    Neu1 and Neu3 on sperm and change during capacitation. A , fluorescent staining of Neu1 on mouse sperm head before and after capacitation and counterstained with DAPI and an unspecific IgG control. The change was quantified using FACS and could be partially inhibited by DANA. B , fluorescent staining of Neu3 on mouse sperm head before and after capacitation and counterstained with DAPI and an unspecific IgG control. The change was quantified using FACS and could be partially inhibited by DANA. A and B are representative of 12 experiments each. C , Neu1/3 on mouse sperm in the testes and the proximal and distal epididymis. Arrowheads show positive staining only in mature sperm of the cauda (distal epididymis). HE , hematoxylin and eosin. D , NEU1/3 on human ejaculated sperm. Expression was highest in the hind part of the sperm head. The counterstain is DAPI ( blue ). ALF488 , Alexa Fluor 488. E , release of Neu1 and Neu3 from sperm into the supernatant and the effect of the sialidase inhibitor DANA. U , uncapacitated; C , capacitated; C + I , capacitated with the inhibitor DANA. F , sialidase activity in and release from sperm are both affected by protein inhibitor ( PI ). ***, p ≤ 0.001 versus the control.

    Journal: The Journal of Biological Chemistry

    Article Title: Sialidases on Mammalian Sperm Mediate Deciduous Sialylation during Capacitation *

    doi: 10.1074/jbc.M112.380584

    Figure Lengend Snippet: Neu1 and Neu3 on sperm and change during capacitation. A , fluorescent staining of Neu1 on mouse sperm head before and after capacitation and counterstained with DAPI and an unspecific IgG control. The change was quantified using FACS and could be partially inhibited by DANA. B , fluorescent staining of Neu3 on mouse sperm head before and after capacitation and counterstained with DAPI and an unspecific IgG control. The change was quantified using FACS and could be partially inhibited by DANA. A and B are representative of 12 experiments each. C , Neu1/3 on mouse sperm in the testes and the proximal and distal epididymis. Arrowheads show positive staining only in mature sperm of the cauda (distal epididymis). HE , hematoxylin and eosin. D , NEU1/3 on human ejaculated sperm. Expression was highest in the hind part of the sperm head. The counterstain is DAPI ( blue ). ALF488 , Alexa Fluor 488. E , release of Neu1 and Neu3 from sperm into the supernatant and the effect of the sialidase inhibitor DANA. U , uncapacitated; C , capacitated; C + I , capacitated with the inhibitor DANA. F , sialidase activity in and release from sperm are both affected by protein inhibitor ( PI ). ***, p ≤ 0.001 versus the control.

    Article Snippet: Sialidase activity was assayed with and without protease inhibitor (protease inhibitor mixture set III, EDTA-free (1:100), Calbiochem) in the capacitation medium and incubated at 37 °C for 2 h.

    Techniques: Staining, FACS, Expressing, Activity Assay

    Release of Sia monosaccharides during sperm capacitation. A and B , released Sia (monosaccharides) from in vitro capacitated sperm. A , Neu5Gc and Neu5Ac from mouse sperm. B , Neu5Ac from human sperm. C and D , increase in exposed galactose as measured by lectins. C , E. crista-galli lectin ( ECL ; representative of five experiments). D , PNA lectin (representative of five experiments). Yellow circles , galactose; blue square , N -acetylglucosamine; yellow square , N -acetylgalactosamine. E and F , sialidase activity during sperm capacitation. E , sialidase activity of uncapacitated and in vitro capacitated mouse sperm with and without addition of the sialidase inhibitor DANA (representative of eight experiments). 4MU , 4-methylumbelliferyl. F , sialidase activity of uncapacitated and in vitro capacitated human sperm with and without addition of the sialidase inhibitor DANA (representative of eight experiments). G , sialidase activity in female mouse uterine fluid collected during estrous and 1.5 h after mating, capacitated mouse cauda epididymal sperm, and mouse sperm retrieved from the uterus 1.5 h after mating (representative of four experiments). *, p ≤ 0.05; **, p ≤ 0.01; ***, p ≤ 0.001 versus the control.

    Journal: The Journal of Biological Chemistry

    Article Title: Sialidases on Mammalian Sperm Mediate Deciduous Sialylation during Capacitation *

    doi: 10.1074/jbc.M112.380584

    Figure Lengend Snippet: Release of Sia monosaccharides during sperm capacitation. A and B , released Sia (monosaccharides) from in vitro capacitated sperm. A , Neu5Gc and Neu5Ac from mouse sperm. B , Neu5Ac from human sperm. C and D , increase in exposed galactose as measured by lectins. C , E. crista-galli lectin ( ECL ; representative of five experiments). D , PNA lectin (representative of five experiments). Yellow circles , galactose; blue square , N -acetylglucosamine; yellow square , N -acetylgalactosamine. E and F , sialidase activity during sperm capacitation. E , sialidase activity of uncapacitated and in vitro capacitated mouse sperm with and without addition of the sialidase inhibitor DANA (representative of eight experiments). 4MU , 4-methylumbelliferyl. F , sialidase activity of uncapacitated and in vitro capacitated human sperm with and without addition of the sialidase inhibitor DANA (representative of eight experiments). G , sialidase activity in female mouse uterine fluid collected during estrous and 1.5 h after mating, capacitated mouse cauda epididymal sperm, and mouse sperm retrieved from the uterus 1.5 h after mating (representative of four experiments). *, p ≤ 0.05; **, p ≤ 0.01; ***, p ≤ 0.001 versus the control.

    Article Snippet: Sialidase activity was assayed with and without protease inhibitor (protease inhibitor mixture set III, EDTA-free (1:100), Calbiochem) in the capacitation medium and incubated at 37 °C for 2 h.

    Techniques: In Vitro, Activity Assay

    Function of neuraminidase during capacitation. A , zona pellucida ( ZP ) binding by uncapacitated sperm ( U ), capacitated sperm ( C ), and sperm capacitated in the presence of the sialidase inhibitor DANA ( C + I ). B , Western blot for phospho-ERK of mouse sperm. C , FACS histogram of mouse sperm stained for PY20. D , Western blot for phospho-ERK and PY20 of human sperm ( C + Sia ) capacitated in the presence of free Neu5Ac. **, p ≤ 0.01 versus the control.

    Journal: The Journal of Biological Chemistry

    Article Title: Sialidases on Mammalian Sperm Mediate Deciduous Sialylation during Capacitation *

    doi: 10.1074/jbc.M112.380584

    Figure Lengend Snippet: Function of neuraminidase during capacitation. A , zona pellucida ( ZP ) binding by uncapacitated sperm ( U ), capacitated sperm ( C ), and sperm capacitated in the presence of the sialidase inhibitor DANA ( C + I ). B , Western blot for phospho-ERK of mouse sperm. C , FACS histogram of mouse sperm stained for PY20. D , Western blot for phospho-ERK and PY20 of human sperm ( C + Sia ) capacitated in the presence of free Neu5Ac. **, p ≤ 0.01 versus the control.

    Article Snippet: Sialidase activity was assayed with and without protease inhibitor (protease inhibitor mixture set III, EDTA-free (1:100), Calbiochem) in the capacitation medium and incubated at 37 °C for 2 h.

    Techniques: Binding Assay, Western Blot, FACS, Staining

    Sialidase activity of T. forsythia NanH and SiaHI expressed in E. coli . Strains KCL116 ( E. coli pET30; lanes 1 and 2), KCL117 [ E. coli BL21(DE3)/pET30:: nanH ; lanes 3 and 4], and KCL120 [ E. coli BL21(DE3)/pET30:: siaHI ; lanes 5 and 6] were grown in LB until

    Journal: Journal of Bacteriology

    Article Title: An Orthologue of Bacteroides fragilis NanH Is the Principal Sialidase in Tannerella forsythia ▿

    doi: 10.1128/JB.01618-08

    Figure Lengend Snippet: Sialidase activity of T. forsythia NanH and SiaHI expressed in E. coli . Strains KCL116 ( E. coli pET30; lanes 1 and 2), KCL117 [ E. coli BL21(DE3)/pET30:: nanH ; lanes 3 and 4], and KCL120 [ E. coli BL21(DE3)/pET30:: siaHI ; lanes 5 and 6] were grown in LB until

    Article Snippet: Quantitative sialidase assays involved measuring 4-methylumbelliferone (4-MU) fluorescence, released from 4-MU-NeuNAc by sialidase activity, in a Fluoroskan Ascent FL fluorimeter (Labsystems Thermo, United Kingdom) with an excitation wavelength of 380 nm and an emission wavelength of 460 nm ( , ).

    Techniques: Activity Assay

    Human vaginal microbial communities-collection and amplification. Step 1 - Anaerobic and aerobic vaginal swabs were collected on the same day from each participant. Aerobic swabs were eluted in sodium acetate buffer (pH 5.5) and sialidase activity was checked in the swab eluates using fluorogenic 4MU-Neu5Ac substrate. Anaerobic swabs were eluted in 2X NYCIII media (in an anaerobic chamber) and the communities were “fresh frozen”, without any amplification / overnight culture , by mixing with cryoprotectant and storing at −80°C. Step 2 - On the day of the experiment - fresh frozen anaerobic vaginal communities, from women who had detectable sialidase activity in their aerobic swab eluates, were thawed at 4°C and diluted 4-fold in NYCIII media (in an anaerobic chamber). The diluted communities were used for co-culture experiments with F. nucleatum .

    Journal: bioRxiv

    Article Title: Glycan cross-feeding drives mutualism between Fusobacterium and the vaginal microbiota

    doi: 10.1101/463349

    Figure Lengend Snippet: Human vaginal microbial communities-collection and amplification. Step 1 - Anaerobic and aerobic vaginal swabs were collected on the same day from each participant. Aerobic swabs were eluted in sodium acetate buffer (pH 5.5) and sialidase activity was checked in the swab eluates using fluorogenic 4MU-Neu5Ac substrate. Anaerobic swabs were eluted in 2X NYCIII media (in an anaerobic chamber) and the communities were “fresh frozen”, without any amplification / overnight culture , by mixing with cryoprotectant and storing at −80°C. Step 2 - On the day of the experiment - fresh frozen anaerobic vaginal communities, from women who had detectable sialidase activity in their aerobic swab eluates, were thawed at 4°C and diluted 4-fold in NYCIII media (in an anaerobic chamber). The diluted communities were used for co-culture experiments with F. nucleatum .

    Article Snippet: The finding that Envigo mice had sialidase activity in their vaginas, but Jackson mice did not, led us to wonder whether the vaginal microbiomes differed in the two groups of mice.

    Techniques: Amplification, Activity Assay, Co-Culture Assay

    Vaginal sialidase activity in E. coli and F. nucleatum inoculated mice, and in vaginal microbiota pools of Jackson and Envigo mice, related to Figure 6 (A-B) Sialidase activity in vaginal washes from individual animals purchased from Envigo, estrogenized, and inoculated with E. coli , or F. nucleatum from 1 to 8 dpi. (C) Sialidase activity (after estrogenization) in vaginal wash pooled specimens (1 microbiota pool = 1 cage = 5 mice) from Jackson and Envigo mice, cultured with or without F. nucleatum (WT or ΩsiaT ). No sialidase activity was observed in cultured vaginal microbial communities from Jackson mice, both with and without F. nucleatum . Data shown here is combined from 2 independent experiments.

    Journal: bioRxiv

    Article Title: Glycan cross-feeding drives mutualism between Fusobacterium and the vaginal microbiota

    doi: 10.1101/463349

    Figure Lengend Snippet: Vaginal sialidase activity in E. coli and F. nucleatum inoculated mice, and in vaginal microbiota pools of Jackson and Envigo mice, related to Figure 6 (A-B) Sialidase activity in vaginal washes from individual animals purchased from Envigo, estrogenized, and inoculated with E. coli , or F. nucleatum from 1 to 8 dpi. (C) Sialidase activity (after estrogenization) in vaginal wash pooled specimens (1 microbiota pool = 1 cage = 5 mice) from Jackson and Envigo mice, cultured with or without F. nucleatum (WT or ΩsiaT ). No sialidase activity was observed in cultured vaginal microbial communities from Jackson mice, both with and without F. nucleatum . Data shown here is combined from 2 independent experiments.

    Article Snippet: The finding that Envigo mice had sialidase activity in their vaginas, but Jackson mice did not, led us to wonder whether the vaginal microbiomes differed in the two groups of mice.

    Techniques: Activity Assay, Mouse Assay, Cell Culture

    F. nucleatum 23726 takes up and catabolizes free sialic acid released by exogenous sialidases Sialidase-producers in the vaginal microbial community release free sialic acids (red diamonds) from host glyco-conjugates, which may be accessed by F. nucleatum , which does not produce sialidase. SiaT = sialic acid transporter; NanA = N -acetylneuraminate lyase; More information about F. nucleatum genes shown in sialic acid catabolic gene cluster, and the enzymes they encode, can be found in Table S2 .

    Journal: bioRxiv

    Article Title: Glycan cross-feeding drives mutualism between Fusobacterium and the vaginal microbiota

    doi: 10.1101/463349

    Figure Lengend Snippet: F. nucleatum 23726 takes up and catabolizes free sialic acid released by exogenous sialidases Sialidase-producers in the vaginal microbial community release free sialic acids (red diamonds) from host glyco-conjugates, which may be accessed by F. nucleatum , which does not produce sialidase. SiaT = sialic acid transporter; NanA = N -acetylneuraminate lyase; More information about F. nucleatum genes shown in sialic acid catabolic gene cluster, and the enzymes they encode, can be found in Table S2 .

    Article Snippet: The finding that Envigo mice had sialidase activity in their vaginas, but Jackson mice did not, led us to wonder whether the vaginal microbiomes differed in the two groups of mice.

    Techniques:

    C57BL/6 mice from Envigo have endogenous vaginal sialidase activity (A) Sialidase activity in vaginal washes of mice (not estrogenized) from different vendors measured using the fluorogenic 4MU-Neu5Ac substrate. N = 10 mice/ vendor. (B) Vaginal sialidase activity is elevated at 72 h post-estrogenization in mice from Envigo. No sialidase activity was detected in vaginal washes of mice from Jackson. N = 40 mice/ vendor. Wilcoxon paired-sign rank test was used for pairwise comparison of sialidase activity before and after estrogen treatment. ** P

    Journal: bioRxiv

    Article Title: Glycan cross-feeding drives mutualism between Fusobacterium and the vaginal microbiota

    doi: 10.1101/463349

    Figure Lengend Snippet: C57BL/6 mice from Envigo have endogenous vaginal sialidase activity (A) Sialidase activity in vaginal washes of mice (not estrogenized) from different vendors measured using the fluorogenic 4MU-Neu5Ac substrate. N = 10 mice/ vendor. (B) Vaginal sialidase activity is elevated at 72 h post-estrogenization in mice from Envigo. No sialidase activity was detected in vaginal washes of mice from Jackson. N = 40 mice/ vendor. Wilcoxon paired-sign rank test was used for pairwise comparison of sialidase activity before and after estrogen treatment. ** P

    Article Snippet: The finding that Envigo mice had sialidase activity in their vaginas, but Jackson mice did not, led us to wonder whether the vaginal microbiomes differed in the two groups of mice.

    Techniques: Mouse Assay, Activity Assay

    Sialic acid catabolism facilitates persistent F. nucleatum vaginal colonization in the setting of an endogenous sialidase-producing microbiome (A) Heat map shows abundance of OTUs in uncultured pooled vaginal specimens (1 pool = 1 cage = 5 mice) collected 1-day post inoculation with F. nucleatum . OTUs were assigned using the UPARSE-OTU algorithm. Data was clustered using hierarchical Euclidean clustering. (B, C) F. nucleatum titers in vaginal wash collected at indicated time points post inoculation (B) in mice from Envigo, and (C) in mice from Jackson. ** P

    Journal: bioRxiv

    Article Title: Glycan cross-feeding drives mutualism between Fusobacterium and the vaginal microbiota

    doi: 10.1101/463349

    Figure Lengend Snippet: Sialic acid catabolism facilitates persistent F. nucleatum vaginal colonization in the setting of an endogenous sialidase-producing microbiome (A) Heat map shows abundance of OTUs in uncultured pooled vaginal specimens (1 pool = 1 cage = 5 mice) collected 1-day post inoculation with F. nucleatum . OTUs were assigned using the UPARSE-OTU algorithm. Data was clustered using hierarchical Euclidean clustering. (B, C) F. nucleatum titers in vaginal wash collected at indicated time points post inoculation (B) in mice from Envigo, and (C) in mice from Jackson. ** P

    Article Snippet: The finding that Envigo mice had sialidase activity in their vaginas, but Jackson mice did not, led us to wonder whether the vaginal microbiomes differed in the two groups of mice.

    Techniques: Mouse Assay

    F. nucleatum can consume free sialic acid in the environment, but can only access sialic acids within glycans if sialidases from other bacteria are present (A-B) E. coli MG1655 ΔnanA was complemented with empty vector, E. coli nanA or putative nanA from F. nucleatum 23726. F.n. = F. nucleatum ; E.c. = E. coli . (A) Growth was assessed in minimal media with sialic acid by measuring absorbance at 600 nm. (B) Lysates of E. coli nanA mutant with empty vector or the complemented strain were incubated with Neu5Ac and its disappearance was monitored over time by DMB-HPLC. (C) Concentration of sialic acid (Neu5Ac) remaining in the medium 24 h post inoculation. Sialic acid consumption by F. nucleatum 23726 wild-type (WT) and ΩsiaT was studied in complete growth medium supplemented with free sialic acid. Data shown is representative of two independent experiments. Error bars show standard deviation from the mean value. (D) Growth of F. nucleatum ( F. nuc.) WT and ΩsiaT in low carbohydrate (carb) medium with or without supplementation with the indicated carbohydrates. Data shown is representative of two independent experiments. (E) Cell associated sialidase activity of anaerobically cultured G. vaginalis JCP8151B and F. nucleatum ATCC23726 strains was analyzed using fluorogenic 4MU-Neu5Ac substrate. (F) F. nucleatum was grown anaerobically in media that was either untreated or exposed to purified sialidase from Arthrobacter ureafaciens ( A.u.) or G. vaginalis ( G.v.). Total and free sialic acid content was measured at 0 and 48 h post inoculation. Bound sialic acids (Bound = Total - Free) are inaccessible to F. nucleatum , except in the presence of exogenous sialidase. Data shown are representative of at least two technical replicates per condition performed in two or more independent experiments. Error bars represent standard deviation from the mean value. See also Figure S1 , S2 and S3 .

    Journal: bioRxiv

    Article Title: Glycan cross-feeding drives mutualism between Fusobacterium and the vaginal microbiota

    doi: 10.1101/463349

    Figure Lengend Snippet: F. nucleatum can consume free sialic acid in the environment, but can only access sialic acids within glycans if sialidases from other bacteria are present (A-B) E. coli MG1655 ΔnanA was complemented with empty vector, E. coli nanA or putative nanA from F. nucleatum 23726. F.n. = F. nucleatum ; E.c. = E. coli . (A) Growth was assessed in minimal media with sialic acid by measuring absorbance at 600 nm. (B) Lysates of E. coli nanA mutant with empty vector or the complemented strain were incubated with Neu5Ac and its disappearance was monitored over time by DMB-HPLC. (C) Concentration of sialic acid (Neu5Ac) remaining in the medium 24 h post inoculation. Sialic acid consumption by F. nucleatum 23726 wild-type (WT) and ΩsiaT was studied in complete growth medium supplemented with free sialic acid. Data shown is representative of two independent experiments. Error bars show standard deviation from the mean value. (D) Growth of F. nucleatum ( F. nuc.) WT and ΩsiaT in low carbohydrate (carb) medium with or without supplementation with the indicated carbohydrates. Data shown is representative of two independent experiments. (E) Cell associated sialidase activity of anaerobically cultured G. vaginalis JCP8151B and F. nucleatum ATCC23726 strains was analyzed using fluorogenic 4MU-Neu5Ac substrate. (F) F. nucleatum was grown anaerobically in media that was either untreated or exposed to purified sialidase from Arthrobacter ureafaciens ( A.u.) or G. vaginalis ( G.v.). Total and free sialic acid content was measured at 0 and 48 h post inoculation. Bound sialic acids (Bound = Total - Free) are inaccessible to F. nucleatum , except in the presence of exogenous sialidase. Data shown are representative of at least two technical replicates per condition performed in two or more independent experiments. Error bars represent standard deviation from the mean value. See also Figure S1 , S2 and S3 .

    Article Snippet: The finding that Envigo mice had sialidase activity in their vaginas, but Jackson mice did not, led us to wonder whether the vaginal microbiomes differed in the two groups of mice.

    Techniques: Plasmid Preparation, Mutagenesis, Incubation, High Performance Liquid Chromatography, Concentration Assay, Standard Deviation, Activity Assay, Cell Culture, Purification

    F. nucleatum ATCC23726 uptakes free sialic acid released by P. bivia sialidases, related to Figure 1 Relative amount of bound (bound = total-free) and free sialic acid remaining in the medium at 0 and 48 hours post inoculation. F. nucleatum was grown anaerobically in media that was either untreated or exposed to purified sialidase from Arthrobacter ureafaciens ( A.u. ) or P. bivia (P.b.) ATCC 29303. Bound sialic acids are inaccessible to F. nucleatum except in the presence of exogenous sialidase. Error bars represent standard deviation of the mean value.

    Journal: bioRxiv

    Article Title: Glycan cross-feeding drives mutualism between Fusobacterium and the vaginal microbiota

    doi: 10.1101/463349

    Figure Lengend Snippet: F. nucleatum ATCC23726 uptakes free sialic acid released by P. bivia sialidases, related to Figure 1 Relative amount of bound (bound = total-free) and free sialic acid remaining in the medium at 0 and 48 hours post inoculation. F. nucleatum was grown anaerobically in media that was either untreated or exposed to purified sialidase from Arthrobacter ureafaciens ( A.u. ) or P. bivia (P.b.) ATCC 29303. Bound sialic acids are inaccessible to F. nucleatum except in the presence of exogenous sialidase. Error bars represent standard deviation of the mean value.

    Article Snippet: The finding that Envigo mice had sialidase activity in their vaginas, but Jackson mice did not, led us to wonder whether the vaginal microbiomes differed in the two groups of mice.

    Techniques: Purification, Standard Deviation

    Sialidase activity in ex vivo cultured vaginal microbial communities from Envigo mice increases in presence of F. nucleatum (A) Sialidase activity in vaginal washes from 1 to 8 dpi from individual animals purchased from Envigo, estrogenized, and inoculated with either WT or ΩsiaT F. nucleatum . Data at later time points were compared to day 1 values using Friedman test, along with correction for multiple planned comparisons using Dunn’s test. (B) Same experiment and data as shown in A but analyzed to compare between WT or ΩsiaT- inoculated animals at each time point using the Mann-Whitney test. Data in A and B represent combined data from two independent experiments. Data points with negative values were set to 0.001 to represent them on the log scale. (C) Sialidase activity in microbiota pools from Envigo mice. Communities were cultured in the presence or absence of F. nucleatum ( F. nuc. ) WT or ΩsiaT . Each “microbiota pool” consists of a cultured vaginal community from pooled vaginal wash of 4-5 co-housed mice. Wilcoxon paired-sign rank test was used for pairwise comparison of sialidase activity in each cultured microbiome compared to the identical microbiome cultured in the presence of F. nucleatum . Data shown is combined from 4 independent biological replicates with 7-8 technical replicates for each microbiota pool. (D) Same experimental data as in C . Data shown is combined from all microbiota pools for each group and analyzed to allow for comparison of the relative sialidase boost between no added F. nucleatum ( F.n. ) versus addition of WT or ΩsiaT . A statistical comparison between the two groups was performed using Wilcoxon matched-pairs signed rank test. Data shown is combined from 3 independent biological replicates. Line in the bar indicates mean value. On all graphs *P

    Journal: bioRxiv

    Article Title: Glycan cross-feeding drives mutualism between Fusobacterium and the vaginal microbiota

    doi: 10.1101/463349

    Figure Lengend Snippet: Sialidase activity in ex vivo cultured vaginal microbial communities from Envigo mice increases in presence of F. nucleatum (A) Sialidase activity in vaginal washes from 1 to 8 dpi from individual animals purchased from Envigo, estrogenized, and inoculated with either WT or ΩsiaT F. nucleatum . Data at later time points were compared to day 1 values using Friedman test, along with correction for multiple planned comparisons using Dunn’s test. (B) Same experiment and data as shown in A but analyzed to compare between WT or ΩsiaT- inoculated animals at each time point using the Mann-Whitney test. Data in A and B represent combined data from two independent experiments. Data points with negative values were set to 0.001 to represent them on the log scale. (C) Sialidase activity in microbiota pools from Envigo mice. Communities were cultured in the presence or absence of F. nucleatum ( F. nuc. ) WT or ΩsiaT . Each “microbiota pool” consists of a cultured vaginal community from pooled vaginal wash of 4-5 co-housed mice. Wilcoxon paired-sign rank test was used for pairwise comparison of sialidase activity in each cultured microbiome compared to the identical microbiome cultured in the presence of F. nucleatum . Data shown is combined from 4 independent biological replicates with 7-8 technical replicates for each microbiota pool. (D) Same experimental data as in C . Data shown is combined from all microbiota pools for each group and analyzed to allow for comparison of the relative sialidase boost between no added F. nucleatum ( F.n. ) versus addition of WT or ΩsiaT . A statistical comparison between the two groups was performed using Wilcoxon matched-pairs signed rank test. Data shown is combined from 3 independent biological replicates. Line in the bar indicates mean value. On all graphs *P

    Article Snippet: The finding that Envigo mice had sialidase activity in their vaginas, but Jackson mice did not, led us to wonder whether the vaginal microbiomes differed in the two groups of mice.

    Techniques: Activity Assay, Ex Vivo, Cell Culture, Mouse Assay, MANN-WHITNEY

    F. nucleatum supports growth and sialidase production by human BV bacteria (A) Human vaginal communities were cultivated anaerobically in Columbia media in the presence or absence of added F. nucleatum . Sialidase activity was measured following anaerobic culture. Communities from 21 individual women were used. Data are combined from 2 independent experiments. A statistical comparison between the two groups was performed using Wilcoxon matched-pairs signed rank test. Negative values were set to 0.0018 (lowest positive value) to depict them on the log scale. (B-C) G. vaginalis ( G.v. ) was co-cultivated anaerobically in Columbia media in the presence or absence of F. nucleatum ( F.n. ), followed by measurement of sialidase activity (B) and viable titers of G. vaginalis (colony forming units, C). Note that G. vaginalis was not detectable under these conditions in the absence of F. nucleatum. In this case, G. vaginalis levels were plotted at one half the limit of detection (LOD=200 CFU/mL). Heat map data is representative of two independent experiments. CFU data is combined from two independent experiments, each with 3 technical replicates each. On all graphs ***P

    Journal: bioRxiv

    Article Title: Glycan cross-feeding drives mutualism between Fusobacterium and the vaginal microbiota

    doi: 10.1101/463349

    Figure Lengend Snippet: F. nucleatum supports growth and sialidase production by human BV bacteria (A) Human vaginal communities were cultivated anaerobically in Columbia media in the presence or absence of added F. nucleatum . Sialidase activity was measured following anaerobic culture. Communities from 21 individual women were used. Data are combined from 2 independent experiments. A statistical comparison between the two groups was performed using Wilcoxon matched-pairs signed rank test. Negative values were set to 0.0018 (lowest positive value) to depict them on the log scale. (B-C) G. vaginalis ( G.v. ) was co-cultivated anaerobically in Columbia media in the presence or absence of F. nucleatum ( F.n. ), followed by measurement of sialidase activity (B) and viable titers of G. vaginalis (colony forming units, C). Note that G. vaginalis was not detectable under these conditions in the absence of F. nucleatum. In this case, G. vaginalis levels were plotted at one half the limit of detection (LOD=200 CFU/mL). Heat map data is representative of two independent experiments. CFU data is combined from two independent experiments, each with 3 technical replicates each. On all graphs ***P

    Article Snippet: The finding that Envigo mice had sialidase activity in their vaginas, but Jackson mice did not, led us to wonder whether the vaginal microbiomes differed in the two groups of mice.

    Techniques: Activity Assay