human foreskin fibroblast hff monolayers  (ATCC)


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    ATCC human foreskin fibroblast hff monolayers
    Human Foreskin Fibroblast Hff Monolayers, supplied by ATCC, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    human foreskin fibroblast hff monolayers  (ATCC)


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    ATCC human foreskin fibroblast hff monolayers
    Human Foreskin Fibroblast Hff Monolayers, supplied by ATCC, 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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    human foreskin fibroblast hff monolayers  (ATCC)


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    ATCC human foreskin fibroblast hff monolayers
    Human Foreskin Fibroblast Hff Monolayers, supplied by ATCC, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    human foreskin fibroblasts hffs monolayers  (ATCC)


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    ATCC human foreskin fibroblasts hffs monolayers
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    human foreskin fibroblasts hffs monolayers  (ATCC)


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    ATCC human foreskin fibroblasts hffs monolayers
    Human Foreskin Fibroblasts Hffs Monolayers, supplied by ATCC, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    human foreskin fibroblast hff monolayers  (ATCC)


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    ATCC human foreskin fibroblast hff monolayers
    GRA12-related proteins localize to tachyzoite dense granules, are secreted into the PV, and behave as a soluble and transmembrane PV proteins. (A) <t>Extracellular</t> <t>tachyzoites</t> were stained with anti-HA (α-HA) and anti-GRA5 (α-GRA5) antibody. Tachyzoite nuclei were stained with 4′,6-diamidino-2-phenylindole (DAPI). Tachyzoites were located using differential interference contrast (DIC) microscopy and imaged by confocal microscopy. Number ( n ) of tachyzoites analyzed ( n = 46 for GRA12A, n = 39 for GRA12B, n = 9 for GRA12C, n = 19 for GRA12D). Bars = 5 μm. (B to D) Human foreskin fibroblast <t>(HFF)</t> cells on coverslips were infected with 0.5 MOI tachyzoites of each strain. Vacuoles were located using DIC microscopy, and parasite and host nuclei were stained with DAPI and imaged by confocal microscopy. Vacuoles were stained with anti-HA and anti-GRA5 (B), anti-GRA2 (C), or anti-GRA1 (D). Number ( n ) of vacuoles analyzed, 6 to 42. Bars = 5 μm. (E) PV proteins were fractionated by ultracentrifugation into high-speed pellet (HSP) and high-speed supernatant (HSS) fractions. Proteins were resolved by electrophoresis, transferred to membranes, and probed with anti-HA rabbit antibody specific for HA-tagged GRA12A, GRA12B, or GRA12D, labeled with goat anti-rabbit IgG coupled to peroxidase, and visualized by chemiluminescence.
    Human Foreskin Fibroblast Hff Monolayers, supplied by ATCC, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "A Family of Toxoplasma gondii Genes Related to GRA12 Regulate Cyst Burdens and Cyst Reactivation"

    Article Title: A Family of Toxoplasma gondii Genes Related to GRA12 Regulate Cyst Burdens and Cyst Reactivation

    Journal: mSphere

    doi: 10.1128/mSphere.00182-21

    GRA12-related proteins localize to tachyzoite dense granules, are secreted into the PV, and behave as a soluble and transmembrane PV proteins. (A) Extracellular tachyzoites were stained with anti-HA (α-HA) and anti-GRA5 (α-GRA5) antibody. Tachyzoite nuclei were stained with 4′,6-diamidino-2-phenylindole (DAPI). Tachyzoites were located using differential interference contrast (DIC) microscopy and imaged by confocal microscopy. Number ( n ) of tachyzoites analyzed ( n = 46 for GRA12A, n = 39 for GRA12B, n = 9 for GRA12C, n = 19 for GRA12D). Bars = 5 μm. (B to D) Human foreskin fibroblast (HFF) cells on coverslips were infected with 0.5 MOI tachyzoites of each strain. Vacuoles were located using DIC microscopy, and parasite and host nuclei were stained with DAPI and imaged by confocal microscopy. Vacuoles were stained with anti-HA and anti-GRA5 (B), anti-GRA2 (C), or anti-GRA1 (D). Number ( n ) of vacuoles analyzed, 6 to 42. Bars = 5 μm. (E) PV proteins were fractionated by ultracentrifugation into high-speed pellet (HSP) and high-speed supernatant (HSS) fractions. Proteins were resolved by electrophoresis, transferred to membranes, and probed with anti-HA rabbit antibody specific for HA-tagged GRA12A, GRA12B, or GRA12D, labeled with goat anti-rabbit IgG coupled to peroxidase, and visualized by chemiluminescence.
    Figure Legend Snippet: GRA12-related proteins localize to tachyzoite dense granules, are secreted into the PV, and behave as a soluble and transmembrane PV proteins. (A) Extracellular tachyzoites were stained with anti-HA (α-HA) and anti-GRA5 (α-GRA5) antibody. Tachyzoite nuclei were stained with 4′,6-diamidino-2-phenylindole (DAPI). Tachyzoites were located using differential interference contrast (DIC) microscopy and imaged by confocal microscopy. Number ( n ) of tachyzoites analyzed ( n = 46 for GRA12A, n = 39 for GRA12B, n = 9 for GRA12C, n = 19 for GRA12D). Bars = 5 μm. (B to D) Human foreskin fibroblast (HFF) cells on coverslips were infected with 0.5 MOI tachyzoites of each strain. Vacuoles were located using DIC microscopy, and parasite and host nuclei were stained with DAPI and imaged by confocal microscopy. Vacuoles were stained with anti-HA and anti-GRA5 (B), anti-GRA2 (C), or anti-GRA1 (D). Number ( n ) of vacuoles analyzed, 6 to 42. Bars = 5 μm. (E) PV proteins were fractionated by ultracentrifugation into high-speed pellet (HSP) and high-speed supernatant (HSS) fractions. Proteins were resolved by electrophoresis, transferred to membranes, and probed with anti-HA rabbit antibody specific for HA-tagged GRA12A, GRA12B, or GRA12D, labeled with goat anti-rabbit IgG coupled to peroxidase, and visualized by chemiluminescence.

    Techniques Used: Staining, Microscopy, Confocal Microscopy, Infection, Electrophoresis, Labeling

    GRA12-related gene knockout mutants exhibit reduced cyst maintenance and cyst reactivation. (A) Reactivation assays were performed using in vitro cysts differentiated for 3 days after HFF cell cultures were infected with 2 × 10 2 tachyzoites of parental PruΔ ku80 , Δ gra12A ::GRA12A or Δ gra12A , Δ gra12B ::GRA12B or Δ gra12B , and Δ gra12D ::GRA12D or Δ gra12D . Percent reactivation (mean ± SEM) from three independent experiments is shown. A one-way ANOVA test revealed significance in reactivation [F(6, 20) = 8.642, P = 0.0001], and a Tukey test was used to calculate P values in comparison to parental control PruΔ ku80 and the respective complemented strain as follows: *, P < 0.05; **, P < 0.01. (B) C57BL/6 female mice ( n = 6 to 13) were infected i.p. with 2 × 10 2 tachyzoites of PruΔ ku80 , Δ gra12B , or Δ gra12B ::GRA12B strain, and brain cyst burdens were measured 3 and 6 weeks postinfection (mean ± SEM). Data shown are cumulative results from two to four independent experiments for each strain. At 3 weeks, PruΔ ku80 (two experiments, n = 8), Δ gra12B (two experiments, n = 8), and GRA12B (four experiments, n = 13) strains are shown. At 6 weeks, PruΔ ku80 (two experiments, n = 6), Δ gra12B (two experiments, n = 6), and GRA12B (three experiments, n = 10) strains are shown. A one-way ANOVA test revealed significance in reactivation [F(5, 45) = 9.909, P < 0.0001], and a Tukey test was used to calculate P values in comparison to parental control PruΔ ku80 and the respective complemented strain as follows: **, P < 0.01; NS, not significant.
    Figure Legend Snippet: GRA12-related gene knockout mutants exhibit reduced cyst maintenance and cyst reactivation. (A) Reactivation assays were performed using in vitro cysts differentiated for 3 days after HFF cell cultures were infected with 2 × 10 2 tachyzoites of parental PruΔ ku80 , Δ gra12A ::GRA12A or Δ gra12A , Δ gra12B ::GRA12B or Δ gra12B , and Δ gra12D ::GRA12D or Δ gra12D . Percent reactivation (mean ± SEM) from three independent experiments is shown. A one-way ANOVA test revealed significance in reactivation [F(6, 20) = 8.642, P = 0.0001], and a Tukey test was used to calculate P values in comparison to parental control PruΔ ku80 and the respective complemented strain as follows: *, P < 0.05; **, P < 0.01. (B) C57BL/6 female mice ( n = 6 to 13) were infected i.p. with 2 × 10 2 tachyzoites of PruΔ ku80 , Δ gra12B , or Δ gra12B ::GRA12B strain, and brain cyst burdens were measured 3 and 6 weeks postinfection (mean ± SEM). Data shown are cumulative results from two to four independent experiments for each strain. At 3 weeks, PruΔ ku80 (two experiments, n = 8), Δ gra12B (two experiments, n = 8), and GRA12B (four experiments, n = 13) strains are shown. At 6 weeks, PruΔ ku80 (two experiments, n = 6), Δ gra12B (two experiments, n = 6), and GRA12B (three experiments, n = 10) strains are shown. A one-way ANOVA test revealed significance in reactivation [F(5, 45) = 9.909, P < 0.0001], and a Tukey test was used to calculate P values in comparison to parental control PruΔ ku80 and the respective complemented strain as follows: **, P < 0.01; NS, not significant.

    Techniques Used: Gene Knockout, In Vitro, Infection

    human foreskin fibroblast hff monolayers  (ATCC)


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    ATCC human foreskin fibroblast hff monolayers
    Human Foreskin Fibroblast Hff Monolayers, supplied by ATCC, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    human foreskin fibroblast hff monolayers  (ATCC)


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    ATCC human foreskin fibroblast hff monolayers
    s-WGA colocalizes with GRA4 at the cyst periphery, and s-WGA accumulation at the cyst periphery is GRA2 independent in immature cysts. PruΔ ku80 - and Δ gra2 -infected <t>human</t> <t>foreskin</t> fibroblast <t>(HFF)</t> cells on coverslips were treated under bradyzoite-inducing conditions to differentiate in vitro cysts for 1 day, 2 days, or 3 days. Cysts were located using DIC microscopy and imaged by confocal microscopy. (A to C) The presence of bradyzoites inside cysts was verified by locating parasite nuclei with DAPI staining (not shown) and verifying that each parasite nucleus was surrounded by the expression of cytosolic GFP (GFP + bradyzoites). Cysts were stained with biotinylated s-WGA and anti-GRA4 antibody. Panels show GFP and s-WGA, s-WGA, anti-GRA4, s-WGA and anti-GRA4, and DIC (cyst walls are indicated by white arrowheads). s-WGA colocalized with GRA4 in 100% of cysts evaluated at day 1 ( n = 11) (A), day 2 ( n = 12) (B), and day 3 ( n = 17) (C). (D to F) Fluorescence intensities of s-WGA and GRA4 were measured at the cyst periphery and within the cyst (cyst interior) in PruΔ ku80 and Δ gra2 cysts at day 1 (D), day 2 (E), and day 3 (F). Data are plotted as the ratio of the mean fluorescence intensity at the cyst periphery to that within the cyst interior ± SEM. The numbers of cysts analyzed for the parental PruΔ ku80 and Δ gra2 strains for s-WGA and GRA4 are as follows: s-WGA ( n = 14, n = 14 cysts) and GRA4 ( n = 11, n = 7 cysts) at day 1 (D), s-WGA ( n = 29, n = 24 cysts) and GRA4 ( n = 12, n = 12 cysts) at day 2 (E), and s-WGA ( n = 36, n = 27 cysts) and GRA4 ( n = 17, n = 16 cysts) at day 3 (F). The numerical ratio for the mean fluorescence intensity is labeled for s-WGA and GRA4. P values were calculated using Student’s t test (*, P < 0.05; **, P < 0.01; NS, not significant).
    Human Foreskin Fibroblast Hff Monolayers, supplied by ATCC, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Succinylated Wheat Germ Agglutinin Colocalizes with the Toxoplasma gondii Cyst Wall Glycoprotein CST1"

    Article Title: Succinylated Wheat Germ Agglutinin Colocalizes with the Toxoplasma gondii Cyst Wall Glycoprotein CST1

    Journal: mSphere

    doi: 10.1128/mSphere.00031-20

    s-WGA colocalizes with GRA4 at the cyst periphery, and s-WGA accumulation at the cyst periphery is GRA2 independent in immature cysts. PruΔ ku80 - and Δ gra2 -infected human foreskin fibroblast (HFF) cells on coverslips were treated under bradyzoite-inducing conditions to differentiate in vitro cysts for 1 day, 2 days, or 3 days. Cysts were located using DIC microscopy and imaged by confocal microscopy. (A to C) The presence of bradyzoites inside cysts was verified by locating parasite nuclei with DAPI staining (not shown) and verifying that each parasite nucleus was surrounded by the expression of cytosolic GFP (GFP + bradyzoites). Cysts were stained with biotinylated s-WGA and anti-GRA4 antibody. Panels show GFP and s-WGA, s-WGA, anti-GRA4, s-WGA and anti-GRA4, and DIC (cyst walls are indicated by white arrowheads). s-WGA colocalized with GRA4 in 100% of cysts evaluated at day 1 ( n = 11) (A), day 2 ( n = 12) (B), and day 3 ( n = 17) (C). (D to F) Fluorescence intensities of s-WGA and GRA4 were measured at the cyst periphery and within the cyst (cyst interior) in PruΔ ku80 and Δ gra2 cysts at day 1 (D), day 2 (E), and day 3 (F). Data are plotted as the ratio of the mean fluorescence intensity at the cyst periphery to that within the cyst interior ± SEM. The numbers of cysts analyzed for the parental PruΔ ku80 and Δ gra2 strains for s-WGA and GRA4 are as follows: s-WGA ( n = 14, n = 14 cysts) and GRA4 ( n = 11, n = 7 cysts) at day 1 (D), s-WGA ( n = 29, n = 24 cysts) and GRA4 ( n = 12, n = 12 cysts) at day 2 (E), and s-WGA ( n = 36, n = 27 cysts) and GRA4 ( n = 17, n = 16 cysts) at day 3 (F). The numerical ratio for the mean fluorescence intensity is labeled for s-WGA and GRA4. P values were calculated using Student’s t test (*, P < 0.05; **, P < 0.01; NS, not significant).
    Figure Legend Snippet: s-WGA colocalizes with GRA4 at the cyst periphery, and s-WGA accumulation at the cyst periphery is GRA2 independent in immature cysts. PruΔ ku80 - and Δ gra2 -infected human foreskin fibroblast (HFF) cells on coverslips were treated under bradyzoite-inducing conditions to differentiate in vitro cysts for 1 day, 2 days, or 3 days. Cysts were located using DIC microscopy and imaged by confocal microscopy. (A to C) The presence of bradyzoites inside cysts was verified by locating parasite nuclei with DAPI staining (not shown) and verifying that each parasite nucleus was surrounded by the expression of cytosolic GFP (GFP + bradyzoites). Cysts were stained with biotinylated s-WGA and anti-GRA4 antibody. Panels show GFP and s-WGA, s-WGA, anti-GRA4, s-WGA and anti-GRA4, and DIC (cyst walls are indicated by white arrowheads). s-WGA colocalized with GRA4 in 100% of cysts evaluated at day 1 ( n = 11) (A), day 2 ( n = 12) (B), and day 3 ( n = 17) (C). (D to F) Fluorescence intensities of s-WGA and GRA4 were measured at the cyst periphery and within the cyst (cyst interior) in PruΔ ku80 and Δ gra2 cysts at day 1 (D), day 2 (E), and day 3 (F). Data are plotted as the ratio of the mean fluorescence intensity at the cyst periphery to that within the cyst interior ± SEM. The numbers of cysts analyzed for the parental PruΔ ku80 and Δ gra2 strains for s-WGA and GRA4 are as follows: s-WGA ( n = 14, n = 14 cysts) and GRA4 ( n = 11, n = 7 cysts) at day 1 (D), s-WGA ( n = 29, n = 24 cysts) and GRA4 ( n = 12, n = 12 cysts) at day 2 (E), and s-WGA ( n = 36, n = 27 cysts) and GRA4 ( n = 17, n = 16 cysts) at day 3 (F). The numerical ratio for the mean fluorescence intensity is labeled for s-WGA and GRA4. P values were calculated using Student’s t test (*, P < 0.05; **, P < 0.01; NS, not significant).

    Techniques Used: Infection, In Vitro, Microscopy, Confocal Microscopy, Staining, Expressing, Fluorescence, Labeling

    human foreskin fibroblast hff monolayers  (ATCC)


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    ATCC human foreskin fibroblast hff monolayers
    Human Foreskin Fibroblast Hff Monolayers, supplied by ATCC, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    human foreskin fibroblast hff monolayers  (ATCC)


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    Human Foreskin Fibroblast Hff Monolayers, supplied by ATCC, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    human foreskin fibroblast hff monolayers  (ATCC)


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    Human Foreskin Fibroblast Hff Monolayers, supplied by ATCC, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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