dolichos biflorus agglutinin dba lectin  (Vector Laboratories)


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    Vector Laboratories dolichos biflorus agglutinin dba lectin
    a , b Multiple small hepatic ducts (yellow arrowheads) visualized by retrograde cholangiography using black ink injection from the fetal duodenum ( a , a’ ), anti-E-cadherin (E-cad) immunostaining ( b ; green fluorescence) and 3D construction of the <t>DBA</t> (Dolichos <t>biflorus</t> <t>agglutinin;</t> bile duct marker) <t>lectin</t> stained serial sections ( b’ ) in Sox17 +/− embryos at embryonic days (E) 18.5. In the wild-type embryo, one large common hepatic duct (CHD) connects to the cystic duct (red arrow). The bar graph in a’ shows the numbers of hilar hepatic ducts per whole liver (mean ± s.e.m; by one-way ANOVA followed by Tukey’s test [wild-type: n = 14, mild Sox17 +/− : n = 4, severe Sox17 +/− : n = 7, maternal: n = 7]). c Schematic representation of Sox17 +/+ and Sox17 +/− mice (biliary atresia [BA] model) in an Alb -cre; ROSA mTmG (Albumin [Alb] – Green Fluorescent Protein [GFP] + ) background (left in c ). Whole mount Alb- GFP + liver (right in c ) and immunostaining ( c’ ) of intrahepatic (green; anti-GFP staining) and extrahepatic (magenta; DBA or anti-E-cad staining) bile duct walls at the hepatic hilus of E18.5 embryos. The ectopic Alb-GFP + cells in hepatic ducts are protruding (white arrowhead) from the hepatic parenchyma (dashed lines). Note the contribution of several GFP + cells in the damaged duct walls in Sox17 +/− embryos (black arrows). The insets show the enlarged view of the area represented by the bracket. CD cystic duct, CBD common bile duct, CHD common hepatic duct, HD hepatic duct, LL left lateral lobe, LM left medial lobe, RM right medial lobe, PV portal vein, GB gallbladder, duo duodenum, Scale bars, 250 μm ( a ), 100 μm ( b ), 25 μm ( c ).
    Dolichos Biflorus Agglutinin Dba Lectin, supplied by Vector Laboratories, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Images

    1) Product Images from "Impact of gallbladder hypoplasia on hilar hepatic ducts in biliary atresia"

    Article Title: Impact of gallbladder hypoplasia on hilar hepatic ducts in biliary atresia

    Journal: Communications Medicine

    doi: 10.1038/s43856-024-00544-5

    a , b Multiple small hepatic ducts (yellow arrowheads) visualized by retrograde cholangiography using black ink injection from the fetal duodenum ( a , a’ ), anti-E-cadherin (E-cad) immunostaining ( b ; green fluorescence) and 3D construction of the DBA (Dolichos biflorus agglutinin; bile duct marker) lectin stained serial sections ( b’ ) in Sox17 +/− embryos at embryonic days (E) 18.5. In the wild-type embryo, one large common hepatic duct (CHD) connects to the cystic duct (red arrow). The bar graph in a’ shows the numbers of hilar hepatic ducts per whole liver (mean ± s.e.m; by one-way ANOVA followed by Tukey’s test [wild-type: n = 14, mild Sox17 +/− : n = 4, severe Sox17 +/− : n = 7, maternal: n = 7]). c Schematic representation of Sox17 +/+ and Sox17 +/− mice (biliary atresia [BA] model) in an Alb -cre; ROSA mTmG (Albumin [Alb] – Green Fluorescent Protein [GFP] + ) background (left in c ). Whole mount Alb- GFP + liver (right in c ) and immunostaining ( c’ ) of intrahepatic (green; anti-GFP staining) and extrahepatic (magenta; DBA or anti-E-cad staining) bile duct walls at the hepatic hilus of E18.5 embryos. The ectopic Alb-GFP + cells in hepatic ducts are protruding (white arrowhead) from the hepatic parenchyma (dashed lines). Note the contribution of several GFP + cells in the damaged duct walls in Sox17 +/− embryos (black arrows). The insets show the enlarged view of the area represented by the bracket. CD cystic duct, CBD common bile duct, CHD common hepatic duct, HD hepatic duct, LL left lateral lobe, LM left medial lobe, RM right medial lobe, PV portal vein, GB gallbladder, duo duodenum, Scale bars, 250 μm ( a ), 100 μm ( b ), 25 μm ( c ).
    Figure Legend Snippet: a , b Multiple small hepatic ducts (yellow arrowheads) visualized by retrograde cholangiography using black ink injection from the fetal duodenum ( a , a’ ), anti-E-cadherin (E-cad) immunostaining ( b ; green fluorescence) and 3D construction of the DBA (Dolichos biflorus agglutinin; bile duct marker) lectin stained serial sections ( b’ ) in Sox17 +/− embryos at embryonic days (E) 18.5. In the wild-type embryo, one large common hepatic duct (CHD) connects to the cystic duct (red arrow). The bar graph in a’ shows the numbers of hilar hepatic ducts per whole liver (mean ± s.e.m; by one-way ANOVA followed by Tukey’s test [wild-type: n = 14, mild Sox17 +/− : n = 4, severe Sox17 +/− : n = 7, maternal: n = 7]). c Schematic representation of Sox17 +/+ and Sox17 +/− mice (biliary atresia [BA] model) in an Alb -cre; ROSA mTmG (Albumin [Alb] – Green Fluorescent Protein [GFP] + ) background (left in c ). Whole mount Alb- GFP + liver (right in c ) and immunostaining ( c’ ) of intrahepatic (green; anti-GFP staining) and extrahepatic (magenta; DBA or anti-E-cad staining) bile duct walls at the hepatic hilus of E18.5 embryos. The ectopic Alb-GFP + cells in hepatic ducts are protruding (white arrowhead) from the hepatic parenchyma (dashed lines). Note the contribution of several GFP + cells in the damaged duct walls in Sox17 +/− embryos (black arrows). The insets show the enlarged view of the area represented by the bracket. CD cystic duct, CBD common bile duct, CHD common hepatic duct, HD hepatic duct, LL left lateral lobe, LM left medial lobe, RM right medial lobe, PV portal vein, GB gallbladder, duo duodenum, Scale bars, 250 μm ( a ), 100 μm ( b ), 25 μm ( c ).

    Techniques Used: Injection, Immunostaining, Fluorescence, Marker, Staining

    a Whole-mount dolichos biflorus agglutinin (DBA; magenta) and anti-alpha-smooth muscle actin (SMA; green)-double staining (upper panels) and anti-SOX17 immunostaining (brown) of sagittal sections of distal sac-like structures (i.e., presumptive gallbladder; lower panels) of Sox17 flox/flox (f/f), Sox17 flox/− (f/−) and Shh -cre; Sox17 flox/− ( Shh- cre ; f /− ) embryos at embryonic days (E) 18.5. The border between gallbladder and cystic duct walls in the Shh -cre; f /− embryos cannot be defined even by anti-SMA staining for gallbladder-specific smooth muscle layers in the presumptive gallbladder region. asterisks, vascular smooth muscle. b Dot plots of the gallbladder (GB) width (i.e., maximum diameter of DBA-positive distal sac-like structure), gallbladder-cystic duct (GB + CD) length and minimum common bile duct (CBD) diameter (y-axis) in three genotypes (i.e., control [f /+ and f/f], heterozygous [f /− and +/−] and homozygous [ Shh- cre ; f /− ] deletion of two Sox17 alleles (x-axis). Note significant reduction in GB width and CBD diameter by Tukey’s honestly significant difference test), in contrast to no change in GB + CD length among three genotypes ( Shh- cre ; f / −: n = 9, f /− or +/−: n = 14, f /+ or f/f: n = 9). c Spearman rank correlation tests between GB width, relative GB width per GB + CD length and minimum CBD diameter in x axis and liver injury level (i.e., serum Alkaline phosphatase [ALP, IU/l] level and degeneration area) on the y-axis (black solid line, p < 0.01; gray solid line, 0.01 ≤ p < 0.05; gray broken line, p ≥ 0.05) ( n = 13). d Schematic illustration of the ripple effects of GB wall hypoplasia on the intrahepatic duct (IHBD) network. Hypoplastic GB wall with reduced SOX17 expression causes reduced GB width, albeit of no change in GB + CD length (left in d ), and it simultaneously causes abnormal formation of a hilar bile duct network (right in d ) as follows: i) deformation of a large common hepatic duct (i.e.; multiple small hepatic ducts); ii) extrahepatic herniation of the IHBD wall; iii) a cloud-like immature IHBD network near the hepatic hilus; and iv) peripheral cholestasis. CBD common bile duct, CD cystic duct, GB gallbladder, IHBD intrahepatic bile duct, HD hepatic duct. Scale bar in a , 100 µm.
    Figure Legend Snippet: a Whole-mount dolichos biflorus agglutinin (DBA; magenta) and anti-alpha-smooth muscle actin (SMA; green)-double staining (upper panels) and anti-SOX17 immunostaining (brown) of sagittal sections of distal sac-like structures (i.e., presumptive gallbladder; lower panels) of Sox17 flox/flox (f/f), Sox17 flox/− (f/−) and Shh -cre; Sox17 flox/− ( Shh- cre ; f /− ) embryos at embryonic days (E) 18.5. The border between gallbladder and cystic duct walls in the Shh -cre; f /− embryos cannot be defined even by anti-SMA staining for gallbladder-specific smooth muscle layers in the presumptive gallbladder region. asterisks, vascular smooth muscle. b Dot plots of the gallbladder (GB) width (i.e., maximum diameter of DBA-positive distal sac-like structure), gallbladder-cystic duct (GB + CD) length and minimum common bile duct (CBD) diameter (y-axis) in three genotypes (i.e., control [f /+ and f/f], heterozygous [f /− and +/−] and homozygous [ Shh- cre ; f /− ] deletion of two Sox17 alleles (x-axis). Note significant reduction in GB width and CBD diameter by Tukey’s honestly significant difference test), in contrast to no change in GB + CD length among three genotypes ( Shh- cre ; f / −: n = 9, f /− or +/−: n = 14, f /+ or f/f: n = 9). c Spearman rank correlation tests between GB width, relative GB width per GB + CD length and minimum CBD diameter in x axis and liver injury level (i.e., serum Alkaline phosphatase [ALP, IU/l] level and degeneration area) on the y-axis (black solid line, p < 0.01; gray solid line, 0.01 ≤ p < 0.05; gray broken line, p ≥ 0.05) ( n = 13). d Schematic illustration of the ripple effects of GB wall hypoplasia on the intrahepatic duct (IHBD) network. Hypoplastic GB wall with reduced SOX17 expression causes reduced GB width, albeit of no change in GB + CD length (left in d ), and it simultaneously causes abnormal formation of a hilar bile duct network (right in d ) as follows: i) deformation of a large common hepatic duct (i.e.; multiple small hepatic ducts); ii) extrahepatic herniation of the IHBD wall; iii) a cloud-like immature IHBD network near the hepatic hilus; and iv) peripheral cholestasis. CBD common bile duct, CD cystic duct, GB gallbladder, IHBD intrahepatic bile duct, HD hepatic duct. Scale bar in a , 100 µm.

    Techniques Used: Double Staining, Immunostaining, Staining, Expressing

    dolichos biflorus agglutinin dba  (Vector Laboratories)


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    Vector Laboratories dolichos biflorus agglutinin dba
    Dolichos Biflorus Agglutinin Dba, supplied by Vector Laboratories, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    dolichos biflorus agglutinin dba  (Vector Laboratories)


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    Vector Laboratories dolichos biflorus agglutinin dba
    Dolichos Biflorus Agglutinin Dba, supplied by Vector Laboratories, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    biotinylated dolichos biflorus agglutinin dba  (Vector Laboratories)


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    Vector Laboratories biotinylated dolichos biflorus agglutinin dba
    Mice were intraperitoneally (i.p.) inoculated with saline (control) or 10,000 WT, IIΔ gra15 , or IIΔ gra15 :GRA15 parasites. Brains and spleens were harvested at 3 weeks post infection (wpi). Mice from these infections were used in Figs 1– . A. Graph of Toxoplasma brain burden as assessed by Q-PCR for the Toxoplasma -specific B1 gene. B. Representative images of a brain tissue cyst stained with Dolichos <t>biflorus</t> <t>agglutinin</t> <t>(DBA).</t> Top image is DBA staining, middle image is staining with anti- Toxoplasma antibodies, and bottom image is merge. C. Quantification of cyst numbers in 8 brain sections per mouse. D. Representative images of Iba1+ cells (microglia/macrophages). Scale bar, 100 μm E . Quantification of the number of Iba-1+ cells. F . Representative images of CD3ε+ cells (T cells). Scale bar, 100 μm. Panels on right are enlarged insert of white box in left panels. G . Quantification of CD3ε+ cells. A, C, E, G . Bars, mean ± SEM. N = 8 fields of view/section, 3 sections/mouse, 5–12 mice/group. For each mouse, the number of cells/section was averaged to create a single point. Data representative of two independent experiments.
    Biotinylated Dolichos Biflorus Agglutinin Dba, supplied by Vector Laboratories, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Toxoplasma type II effector GRA15 has limited influence in vivo"

    Article Title: Toxoplasma type II effector GRA15 has limited influence in vivo

    Journal: PLOS ONE

    doi: 10.1371/journal.pone.0300764

    Mice were intraperitoneally (i.p.) inoculated with saline (control) or 10,000 WT, IIΔ gra15 , or IIΔ gra15 :GRA15 parasites. Brains and spleens were harvested at 3 weeks post infection (wpi). Mice from these infections were used in Figs 1– . A. Graph of Toxoplasma brain burden as assessed by Q-PCR for the Toxoplasma -specific B1 gene. B. Representative images of a brain tissue cyst stained with Dolichos biflorus agglutinin (DBA). Top image is DBA staining, middle image is staining with anti- Toxoplasma antibodies, and bottom image is merge. C. Quantification of cyst numbers in 8 brain sections per mouse. D. Representative images of Iba1+ cells (microglia/macrophages). Scale bar, 100 μm E . Quantification of the number of Iba-1+ cells. F . Representative images of CD3ε+ cells (T cells). Scale bar, 100 μm. Panels on right are enlarged insert of white box in left panels. G . Quantification of CD3ε+ cells. A, C, E, G . Bars, mean ± SEM. N = 8 fields of view/section, 3 sections/mouse, 5–12 mice/group. For each mouse, the number of cells/section was averaged to create a single point. Data representative of two independent experiments.
    Figure Legend Snippet: Mice were intraperitoneally (i.p.) inoculated with saline (control) or 10,000 WT, IIΔ gra15 , or IIΔ gra15 :GRA15 parasites. Brains and spleens were harvested at 3 weeks post infection (wpi). Mice from these infections were used in Figs 1– . A. Graph of Toxoplasma brain burden as assessed by Q-PCR for the Toxoplasma -specific B1 gene. B. Representative images of a brain tissue cyst stained with Dolichos biflorus agglutinin (DBA). Top image is DBA staining, middle image is staining with anti- Toxoplasma antibodies, and bottom image is merge. C. Quantification of cyst numbers in 8 brain sections per mouse. D. Representative images of Iba1+ cells (microglia/macrophages). Scale bar, 100 μm E . Quantification of the number of Iba-1+ cells. F . Representative images of CD3ε+ cells (T cells). Scale bar, 100 μm. Panels on right are enlarged insert of white box in left panels. G . Quantification of CD3ε+ cells. A, C, E, G . Bars, mean ± SEM. N = 8 fields of view/section, 3 sections/mouse, 5–12 mice/group. For each mouse, the number of cells/section was averaged to create a single point. Data representative of two independent experiments.

    Techniques Used: Saline, Infection, Staining

    rhodamine conjugated dolichos biflorus agglutinin dba  (Vector Laboratories)


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    Vector Laboratories rhodamine conjugated dolichos biflorus agglutinin dba
    Rhodamine Conjugated Dolichos Biflorus Agglutinin Dba, supplied by Vector Laboratories, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    fluorescein conjugated dolichos biflorus agglutinin dba  (Vector Laboratories)


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    Vector Laboratories fluorescein conjugated dolichos biflorus agglutinin dba
    (A) HFFs were infected with Tg68 or ME49 tachyzoites at an MOI of 5 for 24, 48, or 72 hr. Cells were fixed and IF staining was performed with anti-SAG1 rabbit antibody and goat anti-rabbit secondary antibody <t>conjugated</t> to Alexa Fluor 647. Hoechst 33342 was used to stain DNA. Vacuoles were visualized using a Cytation 3 Imager and representative images are shown. Scale bar, 50 μm. (B) Violin plot representing diameters of vacuoles of Tg68 or ME49 tachyzoites at indicated timepoints. Data are combined from three independent experiments in which at least 5,000 vacuoles per strain were measured in each experiment. In violin plot, the middle line indicates median, the upper line indicates the 75 th percentile, and the lower line indicates the 25 th percentile. ***, Student’s t-test, P < 0.001. (C) HFFs infected with Tg68 or ME49 tachyzoites were cultured under alkaline pH condition (pH = 8.2, ambient CO 2 ) for 7 days. Cells were fixed and stained with anti-BAG1 mouse mAb 8.25.8 and anti-SAG1 rabbit antibody, followed by goat anti-mouse secondary antibody conjugated to Alexa Fluor 568 and goat anti-rabbit secondary antibody conjugated to Alexa Fluor 647. Dolichos <t>biflorus</t> <t>agglutinin</t> <t>(DBA)</t> conjugated to Alexa Fluor 488 was used to stain the cyst wall and Hoechst 33342 was used to stain DNA. Scale bar, 20 μm. (D) Percentage of DBA-positive BAG1-positive SAG1-negative cysts of Tg68 or ME49. Mean ± SE was plotted for 6 independent experiments, in which at least 100 vacuoles/cysts were counted in each experiment. ***, Student’s t-test, P < 0.001. (E) Neighbor-net analysis of Tg68 and 62 T. gondii isolates based on 804,624 SNPs. Major clades of T. gondii are indicated in different colors. Scale bar, number of SNPs per site.
    Fluorescein Conjugated Dolichos Biflorus Agglutinin Dba, supplied by Vector Laboratories, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Constitutive upregulation of transcription factors underlies permissive bradyzoite differentiation in a natural isolate of Toxoplasma gondii"

    Article Title: Constitutive upregulation of transcription factors underlies permissive bradyzoite differentiation in a natural isolate of Toxoplasma gondii

    Journal: bioRxiv

    doi: 10.1101/2024.02.28.582596

    (A) HFFs were infected with Tg68 or ME49 tachyzoites at an MOI of 5 for 24, 48, or 72 hr. Cells were fixed and IF staining was performed with anti-SAG1 rabbit antibody and goat anti-rabbit secondary antibody conjugated to Alexa Fluor 647. Hoechst 33342 was used to stain DNA. Vacuoles were visualized using a Cytation 3 Imager and representative images are shown. Scale bar, 50 μm. (B) Violin plot representing diameters of vacuoles of Tg68 or ME49 tachyzoites at indicated timepoints. Data are combined from three independent experiments in which at least 5,000 vacuoles per strain were measured in each experiment. In violin plot, the middle line indicates median, the upper line indicates the 75 th percentile, and the lower line indicates the 25 th percentile. ***, Student’s t-test, P < 0.001. (C) HFFs infected with Tg68 or ME49 tachyzoites were cultured under alkaline pH condition (pH = 8.2, ambient CO 2 ) for 7 days. Cells were fixed and stained with anti-BAG1 mouse mAb 8.25.8 and anti-SAG1 rabbit antibody, followed by goat anti-mouse secondary antibody conjugated to Alexa Fluor 568 and goat anti-rabbit secondary antibody conjugated to Alexa Fluor 647. Dolichos biflorus agglutinin (DBA) conjugated to Alexa Fluor 488 was used to stain the cyst wall and Hoechst 33342 was used to stain DNA. Scale bar, 20 μm. (D) Percentage of DBA-positive BAG1-positive SAG1-negative cysts of Tg68 or ME49. Mean ± SE was plotted for 6 independent experiments, in which at least 100 vacuoles/cysts were counted in each experiment. ***, Student’s t-test, P < 0.001. (E) Neighbor-net analysis of Tg68 and 62 T. gondii isolates based on 804,624 SNPs. Major clades of T. gondii are indicated in different colors. Scale bar, number of SNPs per site.
    Figure Legend Snippet: (A) HFFs were infected with Tg68 or ME49 tachyzoites at an MOI of 5 for 24, 48, or 72 hr. Cells were fixed and IF staining was performed with anti-SAG1 rabbit antibody and goat anti-rabbit secondary antibody conjugated to Alexa Fluor 647. Hoechst 33342 was used to stain DNA. Vacuoles were visualized using a Cytation 3 Imager and representative images are shown. Scale bar, 50 μm. (B) Violin plot representing diameters of vacuoles of Tg68 or ME49 tachyzoites at indicated timepoints. Data are combined from three independent experiments in which at least 5,000 vacuoles per strain were measured in each experiment. In violin plot, the middle line indicates median, the upper line indicates the 75 th percentile, and the lower line indicates the 25 th percentile. ***, Student’s t-test, P < 0.001. (C) HFFs infected with Tg68 or ME49 tachyzoites were cultured under alkaline pH condition (pH = 8.2, ambient CO 2 ) for 7 days. Cells were fixed and stained with anti-BAG1 mouse mAb 8.25.8 and anti-SAG1 rabbit antibody, followed by goat anti-mouse secondary antibody conjugated to Alexa Fluor 568 and goat anti-rabbit secondary antibody conjugated to Alexa Fluor 647. Dolichos biflorus agglutinin (DBA) conjugated to Alexa Fluor 488 was used to stain the cyst wall and Hoechst 33342 was used to stain DNA. Scale bar, 20 μm. (D) Percentage of DBA-positive BAG1-positive SAG1-negative cysts of Tg68 or ME49. Mean ± SE was plotted for 6 independent experiments, in which at least 100 vacuoles/cysts were counted in each experiment. ***, Student’s t-test, P < 0.001. (E) Neighbor-net analysis of Tg68 and 62 T. gondii isolates based on 804,624 SNPs. Major clades of T. gondii are indicated in different colors. Scale bar, number of SNPs per site.

    Techniques Used: Infection, Staining, Cell Culture

    biotinylated dolichos biflorus agglutinin dba  (Vector Laboratories)


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    Vector Laboratories biotinylated dolichos biflorus agglutinin dba
    Mice were intraperitoneally (i.p.) inoculated with saline (control) or 10,000 WT, IIΔ gra15 , or IIΔ gra15 :GRA15 parasites. Brains and spleens were harvested at 3 weeks post infection (wpi). Mice from these infections were used in - . A. Graph of Toxoplasma brain burden as assessed by Q-PCR for the Toxoplasma -specific B1 gene. B. Representative images of a brain tissue cyst stained with Dolichos <t>biflorus</t> agglutinin <t>(DBA).</t> Top image is DBA staining, middle image is staining with anti- Toxoplasma antibodies, and bottom image is merge. C. Quantification of cyst numbers in 8 brain sections per mouse. D. Representative images of Iba1+ cells (microglia/macrophages). Scale bar, 100 µm E . Quantification of the number of Iba-1+ cells. F . Representative images of CD3ε+ cells (T cells). Scale bar, 100 µm. Panels on right are enlarged insert of white box in left panels. G . Quantification of CD3ε+ cells. A, C, E, G . Bars, mean ± SEM. N = 8 fields of view/section, 3 sections/mouse, 5-12 mice/group. For each mouse, the number of cells/section was averaged to create a single point. Data representative of two independent experiments.
    Biotinylated Dolichos Biflorus Agglutinin Dba, supplied by Vector Laboratories, 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/biotinylated dolichos biflorus agglutinin dba/product/Vector Laboratories
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    biotinylated dolichos biflorus agglutinin dba - by Bioz Stars, 2024-06
    86/100 stars

    Images

    1) Product Images from "Toxoplasma type II effector GRA15 has limited influence in vivo"

    Article Title: Toxoplasma type II effector GRA15 has limited influence in vivo

    Journal: bioRxiv

    doi: 10.1101/2024.02.23.581829

    Mice were intraperitoneally (i.p.) inoculated with saline (control) or 10,000 WT, IIΔ gra15 , or IIΔ gra15 :GRA15 parasites. Brains and spleens were harvested at 3 weeks post infection (wpi). Mice from these infections were used in - . A. Graph of Toxoplasma brain burden as assessed by Q-PCR for the Toxoplasma -specific B1 gene. B. Representative images of a brain tissue cyst stained with Dolichos biflorus agglutinin (DBA). Top image is DBA staining, middle image is staining with anti- Toxoplasma antibodies, and bottom image is merge. C. Quantification of cyst numbers in 8 brain sections per mouse. D. Representative images of Iba1+ cells (microglia/macrophages). Scale bar, 100 µm E . Quantification of the number of Iba-1+ cells. F . Representative images of CD3ε+ cells (T cells). Scale bar, 100 µm. Panels on right are enlarged insert of white box in left panels. G . Quantification of CD3ε+ cells. A, C, E, G . Bars, mean ± SEM. N = 8 fields of view/section, 3 sections/mouse, 5-12 mice/group. For each mouse, the number of cells/section was averaged to create a single point. Data representative of two independent experiments.
    Figure Legend Snippet: Mice were intraperitoneally (i.p.) inoculated with saline (control) or 10,000 WT, IIΔ gra15 , or IIΔ gra15 :GRA15 parasites. Brains and spleens were harvested at 3 weeks post infection (wpi). Mice from these infections were used in - . A. Graph of Toxoplasma brain burden as assessed by Q-PCR for the Toxoplasma -specific B1 gene. B. Representative images of a brain tissue cyst stained with Dolichos biflorus agglutinin (DBA). Top image is DBA staining, middle image is staining with anti- Toxoplasma antibodies, and bottom image is merge. C. Quantification of cyst numbers in 8 brain sections per mouse. D. Representative images of Iba1+ cells (microglia/macrophages). Scale bar, 100 µm E . Quantification of the number of Iba-1+ cells. F . Representative images of CD3ε+ cells (T cells). Scale bar, 100 µm. Panels on right are enlarged insert of white box in left panels. G . Quantification of CD3ε+ cells. A, C, E, G . Bars, mean ± SEM. N = 8 fields of view/section, 3 sections/mouse, 5-12 mice/group. For each mouse, the number of cells/section was averaged to create a single point. Data representative of two independent experiments.

    Techniques Used: Saline, Infection, Staining

    dolichos biflorus agglutinin dba  (Vector Laboratories)


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    Vector Laboratories dolichos biflorus agglutinin dba
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    dolichos biflorus agglutinin dba  (Vector Laboratories)


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    Vector Laboratories dolichos biflorus agglutinin dba
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    fluorescent tagged dolichos biflorus agglutinin dba  (Vector Laboratories)


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    Vector Laboratories fluorescent tagged dolichos biflorus agglutinin dba
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    Vector Laboratories dolichos biflorus agglutinin dba lectin
    a , b Multiple small hepatic ducts (yellow arrowheads) visualized by retrograde cholangiography using black ink injection from the fetal duodenum ( a , a’ ), anti-E-cadherin (E-cad) immunostaining ( b ; green fluorescence) and 3D construction of the <t>DBA</t> (Dolichos <t>biflorus</t> <t>agglutinin;</t> bile duct marker) <t>lectin</t> stained serial sections ( b’ ) in Sox17 +/− embryos at embryonic days (E) 18.5. In the wild-type embryo, one large common hepatic duct (CHD) connects to the cystic duct (red arrow). The bar graph in a’ shows the numbers of hilar hepatic ducts per whole liver (mean ± s.e.m; by one-way ANOVA followed by Tukey’s test [wild-type: n = 14, mild Sox17 +/− : n = 4, severe Sox17 +/− : n = 7, maternal: n = 7]). c Schematic representation of Sox17 +/+ and Sox17 +/− mice (biliary atresia [BA] model) in an Alb -cre; ROSA mTmG (Albumin [Alb] – Green Fluorescent Protein [GFP] + ) background (left in c ). Whole mount Alb- GFP + liver (right in c ) and immunostaining ( c’ ) of intrahepatic (green; anti-GFP staining) and extrahepatic (magenta; DBA or anti-E-cad staining) bile duct walls at the hepatic hilus of E18.5 embryos. The ectopic Alb-GFP + cells in hepatic ducts are protruding (white arrowhead) from the hepatic parenchyma (dashed lines). Note the contribution of several GFP + cells in the damaged duct walls in Sox17 +/− embryos (black arrows). The insets show the enlarged view of the area represented by the bracket. CD cystic duct, CBD common bile duct, CHD common hepatic duct, HD hepatic duct, LL left lateral lobe, LM left medial lobe, RM right medial lobe, PV portal vein, GB gallbladder, duo duodenum, Scale bars, 250 μm ( a ), 100 μm ( b ), 25 μm ( c ).
    Dolichos Biflorus Agglutinin Dba Lectin, supplied by Vector Laboratories, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Vector Laboratories dolichos biflorus agglutinin dba
    a , b Multiple small hepatic ducts (yellow arrowheads) visualized by retrograde cholangiography using black ink injection from the fetal duodenum ( a , a’ ), anti-E-cadherin (E-cad) immunostaining ( b ; green fluorescence) and 3D construction of the <t>DBA</t> (Dolichos <t>biflorus</t> <t>agglutinin;</t> bile duct marker) <t>lectin</t> stained serial sections ( b’ ) in Sox17 +/− embryos at embryonic days (E) 18.5. In the wild-type embryo, one large common hepatic duct (CHD) connects to the cystic duct (red arrow). The bar graph in a’ shows the numbers of hilar hepatic ducts per whole liver (mean ± s.e.m; by one-way ANOVA followed by Tukey’s test [wild-type: n = 14, mild Sox17 +/− : n = 4, severe Sox17 +/− : n = 7, maternal: n = 7]). c Schematic representation of Sox17 +/+ and Sox17 +/− mice (biliary atresia [BA] model) in an Alb -cre; ROSA mTmG (Albumin [Alb] – Green Fluorescent Protein [GFP] + ) background (left in c ). Whole mount Alb- GFP + liver (right in c ) and immunostaining ( c’ ) of intrahepatic (green; anti-GFP staining) and extrahepatic (magenta; DBA or anti-E-cad staining) bile duct walls at the hepatic hilus of E18.5 embryos. The ectopic Alb-GFP + cells in hepatic ducts are protruding (white arrowhead) from the hepatic parenchyma (dashed lines). Note the contribution of several GFP + cells in the damaged duct walls in Sox17 +/− embryos (black arrows). The insets show the enlarged view of the area represented by the bracket. CD cystic duct, CBD common bile duct, CHD common hepatic duct, HD hepatic duct, LL left lateral lobe, LM left medial lobe, RM right medial lobe, PV portal vein, GB gallbladder, duo duodenum, Scale bars, 250 μm ( a ), 100 μm ( b ), 25 μm ( c ).
    Dolichos Biflorus Agglutinin Dba, supplied by Vector Laboratories, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Vector Laboratories biotinylated dolichos biflorus agglutinin dba
    Mice were intraperitoneally (i.p.) inoculated with saline (control) or 10,000 WT, IIΔ gra15 , or IIΔ gra15 :GRA15 parasites. Brains and spleens were harvested at 3 weeks post infection (wpi). Mice from these infections were used in Figs 1– . A. Graph of Toxoplasma brain burden as assessed by Q-PCR for the Toxoplasma -specific B1 gene. B. Representative images of a brain tissue cyst stained with Dolichos <t>biflorus</t> <t>agglutinin</t> <t>(DBA).</t> Top image is DBA staining, middle image is staining with anti- Toxoplasma antibodies, and bottom image is merge. C. Quantification of cyst numbers in 8 brain sections per mouse. D. Representative images of Iba1+ cells (microglia/macrophages). Scale bar, 100 μm E . Quantification of the number of Iba-1+ cells. F . Representative images of CD3ε+ cells (T cells). Scale bar, 100 μm. Panels on right are enlarged insert of white box in left panels. G . Quantification of CD3ε+ cells. A, C, E, G . Bars, mean ± SEM. N = 8 fields of view/section, 3 sections/mouse, 5–12 mice/group. For each mouse, the number of cells/section was averaged to create a single point. Data representative of two independent experiments.
    Biotinylated Dolichos Biflorus Agglutinin Dba, supplied by Vector Laboratories, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Vector Laboratories rhodamine conjugated dolichos biflorus agglutinin dba
    Mice were intraperitoneally (i.p.) inoculated with saline (control) or 10,000 WT, IIΔ gra15 , or IIΔ gra15 :GRA15 parasites. Brains and spleens were harvested at 3 weeks post infection (wpi). Mice from these infections were used in Figs 1– . A. Graph of Toxoplasma brain burden as assessed by Q-PCR for the Toxoplasma -specific B1 gene. B. Representative images of a brain tissue cyst stained with Dolichos <t>biflorus</t> <t>agglutinin</t> <t>(DBA).</t> Top image is DBA staining, middle image is staining with anti- Toxoplasma antibodies, and bottom image is merge. C. Quantification of cyst numbers in 8 brain sections per mouse. D. Representative images of Iba1+ cells (microglia/macrophages). Scale bar, 100 μm E . Quantification of the number of Iba-1+ cells. F . Representative images of CD3ε+ cells (T cells). Scale bar, 100 μm. Panels on right are enlarged insert of white box in left panels. G . Quantification of CD3ε+ cells. A, C, E, G . Bars, mean ± SEM. N = 8 fields of view/section, 3 sections/mouse, 5–12 mice/group. For each mouse, the number of cells/section was averaged to create a single point. Data representative of two independent experiments.
    Rhodamine Conjugated Dolichos Biflorus Agglutinin Dba, supplied by Vector Laboratories, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Vector Laboratories fluorescein conjugated dolichos biflorus agglutinin dba
    (A) HFFs were infected with Tg68 or ME49 tachyzoites at an MOI of 5 for 24, 48, or 72 hr. Cells were fixed and IF staining was performed with anti-SAG1 rabbit antibody and goat anti-rabbit secondary antibody <t>conjugated</t> to Alexa Fluor 647. Hoechst 33342 was used to stain DNA. Vacuoles were visualized using a Cytation 3 Imager and representative images are shown. Scale bar, 50 μm. (B) Violin plot representing diameters of vacuoles of Tg68 or ME49 tachyzoites at indicated timepoints. Data are combined from three independent experiments in which at least 5,000 vacuoles per strain were measured in each experiment. In violin plot, the middle line indicates median, the upper line indicates the 75 th percentile, and the lower line indicates the 25 th percentile. ***, Student’s t-test, P < 0.001. (C) HFFs infected with Tg68 or ME49 tachyzoites were cultured under alkaline pH condition (pH = 8.2, ambient CO 2 ) for 7 days. Cells were fixed and stained with anti-BAG1 mouse mAb 8.25.8 and anti-SAG1 rabbit antibody, followed by goat anti-mouse secondary antibody conjugated to Alexa Fluor 568 and goat anti-rabbit secondary antibody conjugated to Alexa Fluor 647. Dolichos <t>biflorus</t> <t>agglutinin</t> <t>(DBA)</t> conjugated to Alexa Fluor 488 was used to stain the cyst wall and Hoechst 33342 was used to stain DNA. Scale bar, 20 μm. (D) Percentage of DBA-positive BAG1-positive SAG1-negative cysts of Tg68 or ME49. Mean ± SE was plotted for 6 independent experiments, in which at least 100 vacuoles/cysts were counted in each experiment. ***, Student’s t-test, P < 0.001. (E) Neighbor-net analysis of Tg68 and 62 T. gondii isolates based on 804,624 SNPs. Major clades of T. gondii are indicated in different colors. Scale bar, number of SNPs per site.
    Fluorescein Conjugated Dolichos Biflorus Agglutinin Dba, supplied by Vector Laboratories, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Vector Laboratories fluorescent tagged dolichos biflorus agglutinin dba
    (A) HFFs were infected with Tg68 or ME49 tachyzoites at an MOI of 5 for 24, 48, or 72 hr. Cells were fixed and IF staining was performed with anti-SAG1 rabbit antibody and goat anti-rabbit secondary antibody <t>conjugated</t> to Alexa Fluor 647. Hoechst 33342 was used to stain DNA. Vacuoles were visualized using a Cytation 3 Imager and representative images are shown. Scale bar, 50 μm. (B) Violin plot representing diameters of vacuoles of Tg68 or ME49 tachyzoites at indicated timepoints. Data are combined from three independent experiments in which at least 5,000 vacuoles per strain were measured in each experiment. In violin plot, the middle line indicates median, the upper line indicates the 75 th percentile, and the lower line indicates the 25 th percentile. ***, Student’s t-test, P < 0.001. (C) HFFs infected with Tg68 or ME49 tachyzoites were cultured under alkaline pH condition (pH = 8.2, ambient CO 2 ) for 7 days. Cells were fixed and stained with anti-BAG1 mouse mAb 8.25.8 and anti-SAG1 rabbit antibody, followed by goat anti-mouse secondary antibody conjugated to Alexa Fluor 568 and goat anti-rabbit secondary antibody conjugated to Alexa Fluor 647. Dolichos <t>biflorus</t> <t>agglutinin</t> <t>(DBA)</t> conjugated to Alexa Fluor 488 was used to stain the cyst wall and Hoechst 33342 was used to stain DNA. Scale bar, 20 μm. (D) Percentage of DBA-positive BAG1-positive SAG1-negative cysts of Tg68 or ME49. Mean ± SE was plotted for 6 independent experiments, in which at least 100 vacuoles/cysts were counted in each experiment. ***, Student’s t-test, P < 0.001. (E) Neighbor-net analysis of Tg68 and 62 T. gondii isolates based on 804,624 SNPs. Major clades of T. gondii are indicated in different colors. Scale bar, number of SNPs per site.
    Fluorescent Tagged Dolichos Biflorus Agglutinin Dba, supplied by Vector Laboratories, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    a , b Multiple small hepatic ducts (yellow arrowheads) visualized by retrograde cholangiography using black ink injection from the fetal duodenum ( a , a’ ), anti-E-cadherin (E-cad) immunostaining ( b ; green fluorescence) and 3D construction of the DBA (Dolichos biflorus agglutinin; bile duct marker) lectin stained serial sections ( b’ ) in Sox17 +/− embryos at embryonic days (E) 18.5. In the wild-type embryo, one large common hepatic duct (CHD) connects to the cystic duct (red arrow). The bar graph in a’ shows the numbers of hilar hepatic ducts per whole liver (mean ± s.e.m; by one-way ANOVA followed by Tukey’s test [wild-type: n = 14, mild Sox17 +/− : n = 4, severe Sox17 +/− : n = 7, maternal: n = 7]). c Schematic representation of Sox17 +/+ and Sox17 +/− mice (biliary atresia [BA] model) in an Alb -cre; ROSA mTmG (Albumin [Alb] – Green Fluorescent Protein [GFP] + ) background (left in c ). Whole mount Alb- GFP + liver (right in c ) and immunostaining ( c’ ) of intrahepatic (green; anti-GFP staining) and extrahepatic (magenta; DBA or anti-E-cad staining) bile duct walls at the hepatic hilus of E18.5 embryos. The ectopic Alb-GFP + cells in hepatic ducts are protruding (white arrowhead) from the hepatic parenchyma (dashed lines). Note the contribution of several GFP + cells in the damaged duct walls in Sox17 +/− embryos (black arrows). The insets show the enlarged view of the area represented by the bracket. CD cystic duct, CBD common bile duct, CHD common hepatic duct, HD hepatic duct, LL left lateral lobe, LM left medial lobe, RM right medial lobe, PV portal vein, GB gallbladder, duo duodenum, Scale bars, 250 μm ( a ), 100 μm ( b ), 25 μm ( c ).

    Journal: Communications Medicine

    Article Title: Impact of gallbladder hypoplasia on hilar hepatic ducts in biliary atresia

    doi: 10.1038/s43856-024-00544-5

    Figure Lengend Snippet: a , b Multiple small hepatic ducts (yellow arrowheads) visualized by retrograde cholangiography using black ink injection from the fetal duodenum ( a , a’ ), anti-E-cadherin (E-cad) immunostaining ( b ; green fluorescence) and 3D construction of the DBA (Dolichos biflorus agglutinin; bile duct marker) lectin stained serial sections ( b’ ) in Sox17 +/− embryos at embryonic days (E) 18.5. In the wild-type embryo, one large common hepatic duct (CHD) connects to the cystic duct (red arrow). The bar graph in a’ shows the numbers of hilar hepatic ducts per whole liver (mean ± s.e.m; by one-way ANOVA followed by Tukey’s test [wild-type: n = 14, mild Sox17 +/− : n = 4, severe Sox17 +/− : n = 7, maternal: n = 7]). c Schematic representation of Sox17 +/+ and Sox17 +/− mice (biliary atresia [BA] model) in an Alb -cre; ROSA mTmG (Albumin [Alb] – Green Fluorescent Protein [GFP] + ) background (left in c ). Whole mount Alb- GFP + liver (right in c ) and immunostaining ( c’ ) of intrahepatic (green; anti-GFP staining) and extrahepatic (magenta; DBA or anti-E-cad staining) bile duct walls at the hepatic hilus of E18.5 embryos. The ectopic Alb-GFP + cells in hepatic ducts are protruding (white arrowhead) from the hepatic parenchyma (dashed lines). Note the contribution of several GFP + cells in the damaged duct walls in Sox17 +/− embryos (black arrows). The insets show the enlarged view of the area represented by the bracket. CD cystic duct, CBD common bile duct, CHD common hepatic duct, HD hepatic duct, LL left lateral lobe, LM left medial lobe, RM right medial lobe, PV portal vein, GB gallbladder, duo duodenum, Scale bars, 250 μm ( a ), 100 μm ( b ), 25 μm ( c ).

    Article Snippet: The samples were incubated with rhodamine-labeled dolichos biflorus agglutinin (DBA)-lectin (10 µg/ml; Vector Laboratories, RL-1032) and mouse anti-SMA antibodies (1/100 dilution; Sigma-Aldrich, A5228) for 12 h at 4 °C as previously described .

    Techniques: Injection, Immunostaining, Fluorescence, Marker, Staining

    a Whole-mount dolichos biflorus agglutinin (DBA; magenta) and anti-alpha-smooth muscle actin (SMA; green)-double staining (upper panels) and anti-SOX17 immunostaining (brown) of sagittal sections of distal sac-like structures (i.e., presumptive gallbladder; lower panels) of Sox17 flox/flox (f/f), Sox17 flox/− (f/−) and Shh -cre; Sox17 flox/− ( Shh- cre ; f /− ) embryos at embryonic days (E) 18.5. The border between gallbladder and cystic duct walls in the Shh -cre; f /− embryos cannot be defined even by anti-SMA staining for gallbladder-specific smooth muscle layers in the presumptive gallbladder region. asterisks, vascular smooth muscle. b Dot plots of the gallbladder (GB) width (i.e., maximum diameter of DBA-positive distal sac-like structure), gallbladder-cystic duct (GB + CD) length and minimum common bile duct (CBD) diameter (y-axis) in three genotypes (i.e., control [f /+ and f/f], heterozygous [f /− and +/−] and homozygous [ Shh- cre ; f /− ] deletion of two Sox17 alleles (x-axis). Note significant reduction in GB width and CBD diameter by Tukey’s honestly significant difference test), in contrast to no change in GB + CD length among three genotypes ( Shh- cre ; f / −: n = 9, f /− or +/−: n = 14, f /+ or f/f: n = 9). c Spearman rank correlation tests between GB width, relative GB width per GB + CD length and minimum CBD diameter in x axis and liver injury level (i.e., serum Alkaline phosphatase [ALP, IU/l] level and degeneration area) on the y-axis (black solid line, p < 0.01; gray solid line, 0.01 ≤ p < 0.05; gray broken line, p ≥ 0.05) ( n = 13). d Schematic illustration of the ripple effects of GB wall hypoplasia on the intrahepatic duct (IHBD) network. Hypoplastic GB wall with reduced SOX17 expression causes reduced GB width, albeit of no change in GB + CD length (left in d ), and it simultaneously causes abnormal formation of a hilar bile duct network (right in d ) as follows: i) deformation of a large common hepatic duct (i.e.; multiple small hepatic ducts); ii) extrahepatic herniation of the IHBD wall; iii) a cloud-like immature IHBD network near the hepatic hilus; and iv) peripheral cholestasis. CBD common bile duct, CD cystic duct, GB gallbladder, IHBD intrahepatic bile duct, HD hepatic duct. Scale bar in a , 100 µm.

    Journal: Communications Medicine

    Article Title: Impact of gallbladder hypoplasia on hilar hepatic ducts in biliary atresia

    doi: 10.1038/s43856-024-00544-5

    Figure Lengend Snippet: a Whole-mount dolichos biflorus agglutinin (DBA; magenta) and anti-alpha-smooth muscle actin (SMA; green)-double staining (upper panels) and anti-SOX17 immunostaining (brown) of sagittal sections of distal sac-like structures (i.e., presumptive gallbladder; lower panels) of Sox17 flox/flox (f/f), Sox17 flox/− (f/−) and Shh -cre; Sox17 flox/− ( Shh- cre ; f /− ) embryos at embryonic days (E) 18.5. The border between gallbladder and cystic duct walls in the Shh -cre; f /− embryos cannot be defined even by anti-SMA staining for gallbladder-specific smooth muscle layers in the presumptive gallbladder region. asterisks, vascular smooth muscle. b Dot plots of the gallbladder (GB) width (i.e., maximum diameter of DBA-positive distal sac-like structure), gallbladder-cystic duct (GB + CD) length and minimum common bile duct (CBD) diameter (y-axis) in three genotypes (i.e., control [f /+ and f/f], heterozygous [f /− and +/−] and homozygous [ Shh- cre ; f /− ] deletion of two Sox17 alleles (x-axis). Note significant reduction in GB width and CBD diameter by Tukey’s honestly significant difference test), in contrast to no change in GB + CD length among three genotypes ( Shh- cre ; f / −: n = 9, f /− or +/−: n = 14, f /+ or f/f: n = 9). c Spearman rank correlation tests between GB width, relative GB width per GB + CD length and minimum CBD diameter in x axis and liver injury level (i.e., serum Alkaline phosphatase [ALP, IU/l] level and degeneration area) on the y-axis (black solid line, p < 0.01; gray solid line, 0.01 ≤ p < 0.05; gray broken line, p ≥ 0.05) ( n = 13). d Schematic illustration of the ripple effects of GB wall hypoplasia on the intrahepatic duct (IHBD) network. Hypoplastic GB wall with reduced SOX17 expression causes reduced GB width, albeit of no change in GB + CD length (left in d ), and it simultaneously causes abnormal formation of a hilar bile duct network (right in d ) as follows: i) deformation of a large common hepatic duct (i.e.; multiple small hepatic ducts); ii) extrahepatic herniation of the IHBD wall; iii) a cloud-like immature IHBD network near the hepatic hilus; and iv) peripheral cholestasis. CBD common bile duct, CD cystic duct, GB gallbladder, IHBD intrahepatic bile duct, HD hepatic duct. Scale bar in a , 100 µm.

    Article Snippet: The samples were incubated with rhodamine-labeled dolichos biflorus agglutinin (DBA)-lectin (10 µg/ml; Vector Laboratories, RL-1032) and mouse anti-SMA antibodies (1/100 dilution; Sigma-Aldrich, A5228) for 12 h at 4 °C as previously described .

    Techniques: Double Staining, Immunostaining, Staining, Expressing

    Mice were intraperitoneally (i.p.) inoculated with saline (control) or 10,000 WT, IIΔ gra15 , or IIΔ gra15 :GRA15 parasites. Brains and spleens were harvested at 3 weeks post infection (wpi). Mice from these infections were used in Figs 1– . A. Graph of Toxoplasma brain burden as assessed by Q-PCR for the Toxoplasma -specific B1 gene. B. Representative images of a brain tissue cyst stained with Dolichos biflorus agglutinin (DBA). Top image is DBA staining, middle image is staining with anti- Toxoplasma antibodies, and bottom image is merge. C. Quantification of cyst numbers in 8 brain sections per mouse. D. Representative images of Iba1+ cells (microglia/macrophages). Scale bar, 100 μm E . Quantification of the number of Iba-1+ cells. F . Representative images of CD3ε+ cells (T cells). Scale bar, 100 μm. Panels on right are enlarged insert of white box in left panels. G . Quantification of CD3ε+ cells. A, C, E, G . Bars, mean ± SEM. N = 8 fields of view/section, 3 sections/mouse, 5–12 mice/group. For each mouse, the number of cells/section was averaged to create a single point. Data representative of two independent experiments.

    Journal: PLOS ONE

    Article Title: Toxoplasma type II effector GRA15 has limited influence in vivo

    doi: 10.1371/journal.pone.0300764

    Figure Lengend Snippet: Mice were intraperitoneally (i.p.) inoculated with saline (control) or 10,000 WT, IIΔ gra15 , or IIΔ gra15 :GRA15 parasites. Brains and spleens were harvested at 3 weeks post infection (wpi). Mice from these infections were used in Figs 1– . A. Graph of Toxoplasma brain burden as assessed by Q-PCR for the Toxoplasma -specific B1 gene. B. Representative images of a brain tissue cyst stained with Dolichos biflorus agglutinin (DBA). Top image is DBA staining, middle image is staining with anti- Toxoplasma antibodies, and bottom image is merge. C. Quantification of cyst numbers in 8 brain sections per mouse. D. Representative images of Iba1+ cells (microglia/macrophages). Scale bar, 100 μm E . Quantification of the number of Iba-1+ cells. F . Representative images of CD3ε+ cells (T cells). Scale bar, 100 μm. Panels on right are enlarged insert of white box in left panels. G . Quantification of CD3ε+ cells. A, C, E, G . Bars, mean ± SEM. N = 8 fields of view/section, 3 sections/mouse, 5–12 mice/group. For each mouse, the number of cells/section was averaged to create a single point. Data representative of two independent experiments.

    Article Snippet: These sections were then incubated with biotinylated Dolichos Biflorus Agglutinin (DBA) (Vector laboratories 1031, 1:500) and a polyclonal rabbit anti- Toxoplasma antibody (Thermo Fisher Scientific, PA17252, 1:5000) overnight at 4° C. Samples were then washed and incubated with Streptavidin Cy5 (Life technologies, S21374, 1:500) and goat anti-rabbit 568 secondary (Thermo Fisher Scientific, A11011, 1:500) for 4 hours at room temperature, after which samples were washed to remove residual antibody.

    Techniques: Saline, Infection, Staining

    (A) HFFs were infected with Tg68 or ME49 tachyzoites at an MOI of 5 for 24, 48, or 72 hr. Cells were fixed and IF staining was performed with anti-SAG1 rabbit antibody and goat anti-rabbit secondary antibody conjugated to Alexa Fluor 647. Hoechst 33342 was used to stain DNA. Vacuoles were visualized using a Cytation 3 Imager and representative images are shown. Scale bar, 50 μm. (B) Violin plot representing diameters of vacuoles of Tg68 or ME49 tachyzoites at indicated timepoints. Data are combined from three independent experiments in which at least 5,000 vacuoles per strain were measured in each experiment. In violin plot, the middle line indicates median, the upper line indicates the 75 th percentile, and the lower line indicates the 25 th percentile. ***, Student’s t-test, P < 0.001. (C) HFFs infected with Tg68 or ME49 tachyzoites were cultured under alkaline pH condition (pH = 8.2, ambient CO 2 ) for 7 days. Cells were fixed and stained with anti-BAG1 mouse mAb 8.25.8 and anti-SAG1 rabbit antibody, followed by goat anti-mouse secondary antibody conjugated to Alexa Fluor 568 and goat anti-rabbit secondary antibody conjugated to Alexa Fluor 647. Dolichos biflorus agglutinin (DBA) conjugated to Alexa Fluor 488 was used to stain the cyst wall and Hoechst 33342 was used to stain DNA. Scale bar, 20 μm. (D) Percentage of DBA-positive BAG1-positive SAG1-negative cysts of Tg68 or ME49. Mean ± SE was plotted for 6 independent experiments, in which at least 100 vacuoles/cysts were counted in each experiment. ***, Student’s t-test, P < 0.001. (E) Neighbor-net analysis of Tg68 and 62 T. gondii isolates based on 804,624 SNPs. Major clades of T. gondii are indicated in different colors. Scale bar, number of SNPs per site.

    Journal: bioRxiv

    Article Title: Constitutive upregulation of transcription factors underlies permissive bradyzoite differentiation in a natural isolate of Toxoplasma gondii

    doi: 10.1101/2024.02.28.582596

    Figure Lengend Snippet: (A) HFFs were infected with Tg68 or ME49 tachyzoites at an MOI of 5 for 24, 48, or 72 hr. Cells were fixed and IF staining was performed with anti-SAG1 rabbit antibody and goat anti-rabbit secondary antibody conjugated to Alexa Fluor 647. Hoechst 33342 was used to stain DNA. Vacuoles were visualized using a Cytation 3 Imager and representative images are shown. Scale bar, 50 μm. (B) Violin plot representing diameters of vacuoles of Tg68 or ME49 tachyzoites at indicated timepoints. Data are combined from three independent experiments in which at least 5,000 vacuoles per strain were measured in each experiment. In violin plot, the middle line indicates median, the upper line indicates the 75 th percentile, and the lower line indicates the 25 th percentile. ***, Student’s t-test, P < 0.001. (C) HFFs infected with Tg68 or ME49 tachyzoites were cultured under alkaline pH condition (pH = 8.2, ambient CO 2 ) for 7 days. Cells were fixed and stained with anti-BAG1 mouse mAb 8.25.8 and anti-SAG1 rabbit antibody, followed by goat anti-mouse secondary antibody conjugated to Alexa Fluor 568 and goat anti-rabbit secondary antibody conjugated to Alexa Fluor 647. Dolichos biflorus agglutinin (DBA) conjugated to Alexa Fluor 488 was used to stain the cyst wall and Hoechst 33342 was used to stain DNA. Scale bar, 20 μm. (D) Percentage of DBA-positive BAG1-positive SAG1-negative cysts of Tg68 or ME49. Mean ± SE was plotted for 6 independent experiments, in which at least 100 vacuoles/cysts were counted in each experiment. ***, Student’s t-test, P < 0.001. (E) Neighbor-net analysis of Tg68 and 62 T. gondii isolates based on 804,624 SNPs. Major clades of T. gondii are indicated in different colors. Scale bar, number of SNPs per site.

    Article Snippet: Fluorescein-conjugated Dolichos Biflorus Agglutinin (DBA) was obtained from Vector Laboratories.

    Techniques: Infection, Staining, Cell Culture