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    GE Healthcare ecl prime western blotting detection reagent
    Semi-quantification of ORF2-3 binding to LMH cells. Different concentrations of ORF2-3 were incubated with cells for 1h. Binding was analyzed by Western blotting applying <t>Amersham™</t> <t>ECL™</t> Prime Western Blotting Detection Reagent.
    Ecl Prime Western Blotting Detection Reagent, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 99/100, based on 25251 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/ecl prime western blotting detection reagent/product/GE Healthcare
    Average 99 stars, based on 25251 article reviews
    Price from $9.99 to $1999.99
    ecl prime western blotting detection reagent - by Bioz Stars, 2020-09
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    99
    GE Healthcare amersham ecl prime western blotting detection reagent
    Semi-quantification of ORF2-3 binding to LMH cells. Different concentrations of ORF2-3 were incubated with cells for 1h. Binding was analyzed by Western blotting applying <t>Amersham™</t> <t>ECL™</t> Prime Western Blotting Detection Reagent.
    Amersham Ecl Prime Western Blotting Detection Reagent, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 99/100, based on 86 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/amersham ecl prime western blotting detection reagent/product/GE Healthcare
    Average 99 stars, based on 86 article reviews
    Price from $9.99 to $1999.99
    amersham ecl prime western blotting detection reagent - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    Image Search Results


    Semi-quantification of ORF2-3 binding to LMH cells. Different concentrations of ORF2-3 were incubated with cells for 1h. Binding was analyzed by Western blotting applying Amersham™ ECL™ Prime Western Blotting Detection Reagent.

    Journal: PLoS ONE

    Article Title: C-Terminal Amino Acids 471-507 of Avian Hepatitis E Virus Capsid Protein Are Crucial for Binding to Avian and Human Cells

    doi: 10.1371/journal.pone.0153723

    Figure Lengend Snippet: Semi-quantification of ORF2-3 binding to LMH cells. Different concentrations of ORF2-3 were incubated with cells for 1h. Binding was analyzed by Western blotting applying Amersham™ ECL™ Prime Western Blotting Detection Reagent.

    Article Snippet: Amersham™ ECL™ Prime Western Blotting Detection Reagent (GE Healthcare, Little Chalfont, Bukinghamshire, UK) was applied for semi-quantitative analysis and SuperSignal™ West Pico Chemiluminescent Substrate (Pierce, Thermo Fisher Scientific, Life Technology, Carlsbad, CA, USA) was used for qualitative detection

    Techniques: Binding Assay, Incubation, Western Blot

    Ligand-blotting analysis of the interaction between recombinant polypeptides P40 and P26 with proteins from whole worm extracts. Forty µg of recombinant P40 and P26 were labeled with fluorescein, subjected to SDS-PAGE and photographed under a short-UV lamp (a). Whole worm extract was subjected to SDS-PAGE (T = 10%) and transferred onto Hybond C membranes. Membrane strips containing the fractionated extract were incubated with fluorescein-labeled P40 (50 µg; 1125 pmoles)(b) and P26 (40 µg; 1520 pmoles) (c). After extensive washing, the bound recombinant fluorescein-labeled polypeptides were detected using an anti-fluorescein antibody labeled with peroxidase. The peroxidase activity was detected using chemiluminescence. The chemiluminescence reaction was exposed to Hyperfilm ECL (GE Healthcare Bio-Sciences Corp., Piscataway, NJ, USA) for 30 s in both experiments.

    Journal: PLoS ONE

    Article Title: The Molecular and Structural Characterization of Two Vitellogenins from the Free-Living Nematode Oscheius tipulae

    doi: 10.1371/journal.pone.0053460

    Figure Lengend Snippet: Ligand-blotting analysis of the interaction between recombinant polypeptides P40 and P26 with proteins from whole worm extracts. Forty µg of recombinant P40 and P26 were labeled with fluorescein, subjected to SDS-PAGE and photographed under a short-UV lamp (a). Whole worm extract was subjected to SDS-PAGE (T = 10%) and transferred onto Hybond C membranes. Membrane strips containing the fractionated extract were incubated with fluorescein-labeled P40 (50 µg; 1125 pmoles)(b) and P26 (40 µg; 1520 pmoles) (c). After extensive washing, the bound recombinant fluorescein-labeled polypeptides were detected using an anti-fluorescein antibody labeled with peroxidase. The peroxidase activity was detected using chemiluminescence. The chemiluminescence reaction was exposed to Hyperfilm ECL (GE Healthcare Bio-Sciences Corp., Piscataway, NJ, USA) for 30 s in both experiments.

    Article Snippet: After another three washes in TBS, the proteins were detected using the ECL Plus™ Western Blotting Detection System (GE Healthcare Bio-Sciences Corp., Piscataway, NJ, USA) and exposed to X-ray film at 25°C.

    Techniques: Recombinant, Labeling, SDS Page, Incubation, Activity Assay

    The effect of Cadm1 over-expression on tumor growth and metastasis in vivo . (a) Expression of endogenous and exogenous V5-epitope tagged Cadm1 protein (IB, immunoblot) and total protein by ECL densitometry on immunoblot. (d) Total RNA levels of Cadm1 measured by qRT-PCR (mean +/− standard error of the mean [SEM]). Mvt-1 and 6DT1 mouse mammary tumor cells stably expressing Cadm1 were implanted into mammary fat pads of syngeneic FVB mice. Data on primary tumor burden (Mvt-1: p = 0.0031; 6DT1: p = 0.2261) (b), pulmonary surface metastases (c) was collected on day 30 (Mvt-1: p = 0.0026; 6DT1: p = 0.0089; median +/− interquartile range; n = 10 mice per group). (e) Pulmonary surface metastases were normalized by primary tumor mass to determine if Cadm1 overexpression showed an effect on metastasis independent of its potential tumor suppressive activity (Mvt-1: p = 0.0048; 6DT1: p = 0.0185; median +/− interquartile range; n = 10 mice per group). Arrows indicate several but not all visible metastatic nodules present on lung surfaces. (f) Images of whole lungs of two mice implanted with 6DT1 control cells and two mice implanted with 6DT1 cells expressing Cadm1 . (g) Results of tail vein injection study (Mvt-1: p = 0.0361; 6DT1: p = 0.0475; median +/− interquartile range; n = 10 mice per group). (N.S., not significant; * defined as p

    Journal: PLoS Genetics

    Article Title: Cadm1 Is a Metastasis Susceptibility Gene That Suppresses Metastasis by Modifying Tumor Interaction with the Cell-Mediated ImmunityThe Long Path from QTL to Gene

    doi: 10.1371/journal.pgen.1002926

    Figure Lengend Snippet: The effect of Cadm1 over-expression on tumor growth and metastasis in vivo . (a) Expression of endogenous and exogenous V5-epitope tagged Cadm1 protein (IB, immunoblot) and total protein by ECL densitometry on immunoblot. (d) Total RNA levels of Cadm1 measured by qRT-PCR (mean +/− standard error of the mean [SEM]). Mvt-1 and 6DT1 mouse mammary tumor cells stably expressing Cadm1 were implanted into mammary fat pads of syngeneic FVB mice. Data on primary tumor burden (Mvt-1: p = 0.0031; 6DT1: p = 0.2261) (b), pulmonary surface metastases (c) was collected on day 30 (Mvt-1: p = 0.0026; 6DT1: p = 0.0089; median +/− interquartile range; n = 10 mice per group). (e) Pulmonary surface metastases were normalized by primary tumor mass to determine if Cadm1 overexpression showed an effect on metastasis independent of its potential tumor suppressive activity (Mvt-1: p = 0.0048; 6DT1: p = 0.0185; median +/− interquartile range; n = 10 mice per group). Arrows indicate several but not all visible metastatic nodules present on lung surfaces. (f) Images of whole lungs of two mice implanted with 6DT1 control cells and two mice implanted with 6DT1 cells expressing Cadm1 . (g) Results of tail vein injection study (Mvt-1: p = 0.0361; 6DT1: p = 0.0475; median +/− interquartile range; n = 10 mice per group). (N.S., not significant; * defined as p

    Article Snippet: Immunoblot was visualized using Amersham ECL Prime Western Blotting Detection System and Amersham Hyperfilm ECL (GE Healthcare).

    Techniques: Over Expression, In Vivo, Expressing, Quantitative RT-PCR, Stable Transfection, Mouse Assay, Activity Assay, Injection

    The effect of Cadm1 knockdown on tumor growth and metastasis in vivo . 6DT1 cells expressing shRNA constructs targeting Cadm1 showed (a) protein level knockdown of Cadm1 by immunoblot and ECL densitometry quantification and (d) total RNA level knockdown (mean +/− SEM). Orthotopic implant data showing the effects of Cadm1 knockdown on primary tumor burden (shScr vs. sh14: p = 0.0090; shScr vs. sh15: p = 0.0943; sh14 vs. sh15: p = 0.0001) (b), pulmonary surface metastases (shScr vs. sh14: p = 0.0085; shScr vs. sh15: p = 0.0709; sh14 vs. sh15: p = 0.3485) (c), and tumor-normalized metastases (e) ((shScr vs. sh14: p = 0.00005; shScr vs. sh15: p = 0.0444; sh14 vs. sh15: p = 0.0152; median +/− interquartile range; n = 10 mice per group).

    Journal: PLoS Genetics

    Article Title: Cadm1 Is a Metastasis Susceptibility Gene That Suppresses Metastasis by Modifying Tumor Interaction with the Cell-Mediated ImmunityThe Long Path from QTL to Gene

    doi: 10.1371/journal.pgen.1002926

    Figure Lengend Snippet: The effect of Cadm1 knockdown on tumor growth and metastasis in vivo . 6DT1 cells expressing shRNA constructs targeting Cadm1 showed (a) protein level knockdown of Cadm1 by immunoblot and ECL densitometry quantification and (d) total RNA level knockdown (mean +/− SEM). Orthotopic implant data showing the effects of Cadm1 knockdown on primary tumor burden (shScr vs. sh14: p = 0.0090; shScr vs. sh15: p = 0.0943; sh14 vs. sh15: p = 0.0001) (b), pulmonary surface metastases (shScr vs. sh14: p = 0.0085; shScr vs. sh15: p = 0.0709; sh14 vs. sh15: p = 0.3485) (c), and tumor-normalized metastases (e) ((shScr vs. sh14: p = 0.00005; shScr vs. sh15: p = 0.0444; sh14 vs. sh15: p = 0.0152; median +/− interquartile range; n = 10 mice per group).

    Article Snippet: Immunoblot was visualized using Amersham ECL Prime Western Blotting Detection System and Amersham Hyperfilm ECL (GE Healthcare).

    Techniques: In Vivo, Expressing, shRNA, Construct, Mouse Assay

    Detection of hypo-acetylated K16 on histone H4 in the Jmjd1C gt/gt testis. Approximately 15 μg protein extracts from +/+ and gt/gt testes (postnatal day 56 littermates) were applied on SDS-PAGE (10–20% gradient gel) and blotted onto membranes. A) The membranes were reacted with a mixture of three antibodies against HSC70t, histone H3 (H3) and acetylated H4K16 (H4K16Ac) (left) or four antibodies against VASA, MOF, histone H3 and acetylated H4K16 (right) and then detected with HRP-conjugated anti-IgG and an ECL detection reagent. Specific reactant of each antibody used in this mixed immunoblotting is shown in S5 Fig . B) Comparison of the detected protein bands between the +/+ and gt/gt extracts. C) Semi-quantitative comparison of the acetylated H4K16 bands between the +/+ and gt/gt testes extracts. Values obtained from the +/+ and gt/gt testes were normalized with those of the histone H3, VASA and HSC70t protein bands, which roughly corresponded to the numbers of total cells, germ cells at stages from spermatogonium to late spermatid, and spermatids, respectively. Bars in the graph show relative ratios (%) of values of the H4K16ac band in gt/gt against those in +/+ after normalizing with values of standard protein bands as indicated below. Error bars indicate the SEM (n = 4–6).

    Journal: PLoS ONE

    Article Title: JMJD1C Exhibits Multiple Functions in Epigenetic Regulation during Spermatogenesis

    doi: 10.1371/journal.pone.0163466

    Figure Lengend Snippet: Detection of hypo-acetylated K16 on histone H4 in the Jmjd1C gt/gt testis. Approximately 15 μg protein extracts from +/+ and gt/gt testes (postnatal day 56 littermates) were applied on SDS-PAGE (10–20% gradient gel) and blotted onto membranes. A) The membranes were reacted with a mixture of three antibodies against HSC70t, histone H3 (H3) and acetylated H4K16 (H4K16Ac) (left) or four antibodies against VASA, MOF, histone H3 and acetylated H4K16 (right) and then detected with HRP-conjugated anti-IgG and an ECL detection reagent. Specific reactant of each antibody used in this mixed immunoblotting is shown in S5 Fig . B) Comparison of the detected protein bands between the +/+ and gt/gt extracts. C) Semi-quantitative comparison of the acetylated H4K16 bands between the +/+ and gt/gt testes extracts. Values obtained from the +/+ and gt/gt testes were normalized with those of the histone H3, VASA and HSC70t protein bands, which roughly corresponded to the numbers of total cells, germ cells at stages from spermatogonium to late spermatid, and spermatids, respectively. Bars in the graph show relative ratios (%) of values of the H4K16ac band in gt/gt against those in +/+ after normalizing with values of standard protein bands as indicated below. Error bars indicate the SEM (n = 4–6).

    Article Snippet: The blotted membranes (Immobilon-XP) were immunoreacted with primary antibodies and visualized with the ECL Prime Western blotting Detection kit (GE Healthcare) containing secondary antibodies, HRP (horseradish peroxidase)-labeled anti-mouse IgG and anti-rabbit IgG, and the Quant LAS4000-mini (GE Healthcare) in accordance with the manufacturer’s instruction.

    Techniques: SDS Page

    Irgm3 deficiency reverts the IRG recruitment and clearance defects of both Δ rop18 and Δ rop5 parasites in vitro . (A) Western blot analysis of wild type (RH Δku80 ), ROP18 deficient (RH Δku80Δrop18 ) and ROP18 complemented (RH Δku80Δrop18 Complement) parasites. Western blot probed with rabbit anti-ROP18 and rabbit anti-actin flowed by goat anti-rabbit conjugated to HRP and detected by ECL. (B) Survival of female outbred CD1 mice challenged with 100 parasites by i.p. inoculation. n = 5 animals per group from a single experiment. In vitro clearance of parasites in wild type C57BL/6 (WT) (C) or Irgm3 deficient (Irgm3 −/− ) (D) bone marrow derived macrophages following IFN-γ activation (50 units/mL). Means ± S.D. n = 6 samples from 2 combined experiments. For statistical analysis, parasite survival in Irgm3 deficient cells was compared to survival in wild-type cells. Student's t test, ** P

    Journal: PLoS Pathogens

    Article Title: The Polymorphic Pseudokinase ROP5 Controls Virulence in Toxoplasma gondii by Regulating the Active Kinase ROP18

    doi: 10.1371/journal.ppat.1002992

    Figure Lengend Snippet: Irgm3 deficiency reverts the IRG recruitment and clearance defects of both Δ rop18 and Δ rop5 parasites in vitro . (A) Western blot analysis of wild type (RH Δku80 ), ROP18 deficient (RH Δku80Δrop18 ) and ROP18 complemented (RH Δku80Δrop18 Complement) parasites. Western blot probed with rabbit anti-ROP18 and rabbit anti-actin flowed by goat anti-rabbit conjugated to HRP and detected by ECL. (B) Survival of female outbred CD1 mice challenged with 100 parasites by i.p. inoculation. n = 5 animals per group from a single experiment. In vitro clearance of parasites in wild type C57BL/6 (WT) (C) or Irgm3 deficient (Irgm3 −/− ) (D) bone marrow derived macrophages following IFN-γ activation (50 units/mL). Means ± S.D. n = 6 samples from 2 combined experiments. For statistical analysis, parasite survival in Irgm3 deficient cells was compared to survival in wild-type cells. Student's t test, ** P

    Article Snippet: Blots were washed, incubated with goat anti-rabbit IgG conjugated to HRP (Jackson ImmunoResearch), and detected using the ECL Plus western blotting system (GE Healthcare) and FLA5000 phosphorimager analysis (Fuji Life Sciences).

    Techniques: In Vitro, Western Blot, Mouse Assay, Derivative Assay, Activation Assay

    [ NSI + ] strain contains [ PIN + ] prion which acts as the enhancer of the nonsense suppression. (A) SDD-AGE of protein lysates extracted from the 1-1-D931 [ NSI + ] and 1-1-1-D931 [ nsi - ] strains expressing pCUP1-RNQ1-CFP(LEU2) plasmid. Protein lysates were treated with 1% SDS at room temperature. SDS-resistant aggregates of Rnq1-CFP were detected using monoclonal rabbit primary antibodies against GFP [E385] (ab32146) (Abcam, Great Britain) and ECL Prime Western Blotting Detection Reagent kit (GE Healthcare, USA). (B) The effects of RNQ1 deletion on the [ NSI + ] phenotypic manifestation. RNQ1 deletion was obtained as described in Materials and Methods. The [ nsi - ] and [ nsi - ] rnq Δ strains were obtained from the corresponding [ NSI + ] strains by GuHCl treatment. To express RNQ1 , the 5-1-1-D931 [ NSI + ] rnq Δ strain and its [ nsi - ] derivative were transformed with the YGPM25a02 plasmid containing a genomic fragment encoding RNQ1 under the control of its endogenous promoter. Other strains presented in this Figure were transformed with an empty vector expressing only the LEU2 gene. Transformants were selected on–Leu medium with 150 μM CuSO 4 and replica-plated on–Leu–Ade medium or–Leu medium with 150 μM CuSO 4 containing galactose as the sole carbon source. Images were taken after 5 days of incubation of–Ade plates or after 3 passages on Gal plates.

    Journal: PLoS Genetics

    Article Title: Interaction of Prions Causes Heritable Traits in Saccharomyces cerevisiae

    doi: 10.1371/journal.pgen.1006504

    Figure Lengend Snippet: [ NSI + ] strain contains [ PIN + ] prion which acts as the enhancer of the nonsense suppression. (A) SDD-AGE of protein lysates extracted from the 1-1-D931 [ NSI + ] and 1-1-1-D931 [ nsi - ] strains expressing pCUP1-RNQ1-CFP(LEU2) plasmid. Protein lysates were treated with 1% SDS at room temperature. SDS-resistant aggregates of Rnq1-CFP were detected using monoclonal rabbit primary antibodies against GFP [E385] (ab32146) (Abcam, Great Britain) and ECL Prime Western Blotting Detection Reagent kit (GE Healthcare, USA). (B) The effects of RNQ1 deletion on the [ NSI + ] phenotypic manifestation. RNQ1 deletion was obtained as described in Materials and Methods. The [ nsi - ] and [ nsi - ] rnq Δ strains were obtained from the corresponding [ NSI + ] strains by GuHCl treatment. To express RNQ1 , the 5-1-1-D931 [ NSI + ] rnq Δ strain and its [ nsi - ] derivative were transformed with the YGPM25a02 plasmid containing a genomic fragment encoding RNQ1 under the control of its endogenous promoter. Other strains presented in this Figure were transformed with an empty vector expressing only the LEU2 gene. Transformants were selected on–Leu medium with 150 μM CuSO 4 and replica-plated on–Leu–Ade medium or–Leu medium with 150 μM CuSO 4 containing galactose as the sole carbon source. Images were taken after 5 days of incubation of–Ade plates or after 3 passages on Gal plates.

    Article Snippet: Proteins fused with CFP, GFP, and YFP were detected using monoclonal rabbit primary antibodies against GFP [E385] (ab32146) (Abcam, Great Britain) and the Amersham ECL Prime Western Blotting Detection Reagent kit (GE Healthcare, USA).

    Techniques: Expressing, Plasmid Preparation, Western Blot, Transformation Assay, Incubation

    Mit1 is not a determinant of the [ NSI + ] factor. (A) SDD-AGE assay of protein lysates extracted from the 4-1-1-D931 [ NSI + ] and 1-4-1-1-D931 [ nsi - ] strains expressing pMIT1-MIT1-GFP(URA3) plasmid. Cells were grown for 48 h at 30°C in liquid–Ura selective medium containing 150 μM CuSO 4 . Protein lysates were treated with 1% SDS at room temperature. Mit1-GFP was detected with monoclonal rabbit primary antibodies against GFP [E385] (ab32146) (Abcam, Great Britain) and ECL Prime Western Blotting Detection Reagent kit (GE Healthcare, USA). (B) MIT1 deletion does not affect the [ NSI + ] phenotypic manifestation. MIT1 deletion was obtained as described in Materials and Methods. The [ nsi - ] derivative of the 2–936 [ NSI + ] mit1 Δ was obtained by GuHCl treatment. To express MIT1 , 2–936 [ NSI + ] mit1 Δ strain was transformed with YGPM21o12 plasmid from the YSC4613 genomic library containing a genomic fragment encoding MIT1 under the control of its endogenous promoter. Other strains presented in this Figure were transformed with a vector expressing only the LEU2 gene. Transformants were selected on–Leu medium with 150 μM CuSO 4 and replica-plated on–Leu–Ade or–Leu Gal media with 150 μM CuSO 4 . Images were taken after 5 days of incubation of–Ade plates or after 3 passages on Gal plates.

    Journal: PLoS Genetics

    Article Title: Interaction of Prions Causes Heritable Traits in Saccharomyces cerevisiae

    doi: 10.1371/journal.pgen.1006504

    Figure Lengend Snippet: Mit1 is not a determinant of the [ NSI + ] factor. (A) SDD-AGE assay of protein lysates extracted from the 4-1-1-D931 [ NSI + ] and 1-4-1-1-D931 [ nsi - ] strains expressing pMIT1-MIT1-GFP(URA3) plasmid. Cells were grown for 48 h at 30°C in liquid–Ura selective medium containing 150 μM CuSO 4 . Protein lysates were treated with 1% SDS at room temperature. Mit1-GFP was detected with monoclonal rabbit primary antibodies against GFP [E385] (ab32146) (Abcam, Great Britain) and ECL Prime Western Blotting Detection Reagent kit (GE Healthcare, USA). (B) MIT1 deletion does not affect the [ NSI + ] phenotypic manifestation. MIT1 deletion was obtained as described in Materials and Methods. The [ nsi - ] derivative of the 2–936 [ NSI + ] mit1 Δ was obtained by GuHCl treatment. To express MIT1 , 2–936 [ NSI + ] mit1 Δ strain was transformed with YGPM21o12 plasmid from the YSC4613 genomic library containing a genomic fragment encoding MIT1 under the control of its endogenous promoter. Other strains presented in this Figure were transformed with a vector expressing only the LEU2 gene. Transformants were selected on–Leu medium with 150 μM CuSO 4 and replica-plated on–Leu–Ade or–Leu Gal media with 150 μM CuSO 4 . Images were taken after 5 days of incubation of–Ade plates or after 3 passages on Gal plates.

    Article Snippet: Proteins fused with CFP, GFP, and YFP were detected using monoclonal rabbit primary antibodies against GFP [E385] (ab32146) (Abcam, Great Britain) and the Amersham ECL Prime Western Blotting Detection Reagent kit (GE Healthcare, USA).

    Techniques: Expressing, Plasmid Preparation, Western Blot, Transformation Assay, Incubation

    [ SWI + ] prion is a key determinant of nonsense suppression in [ NSI + ] strains. (A) Sedimentation analysis of Swi1(1–297)-YFP protein from the 4-1-1-D931 [ NSI + ] and 1-4-1-1-D931 [ nsi - ] strains expressing pCUP1-SWI1(1–297)-YFP (URA3) plasmid. Soluble (S) and insoluble (I) fractions were obtained as indicated in Materials and Methods. Swi1(1–297)-YFP was detected using monoclonal rabbit primary antibodies against GFP [E385] (ab32146) (Abcam, Great Britain) and ECL Prime Western Blotting Detection Reagent kit (GE Healthcare, USA). Next, SDD-AGE analysis of insoluble fractions of [ NSI + ] and [ nsi - ] strains comprising Swi1(1–297)-YFP was performed. (B) The effects of SWI1 deletion on the [ NSI + ] phenotypic manifestation. SWI1 deletion was obtained as described in Materials and Methods. To express SWI1 , the 11-1-1-D931 [ NSI + ] swi1 Δ strain was transformed with the YGPM19p21 plasmid from the YSC4613 genomic library, containing a genomic fragment encoding SWI1 under the control of its endogenous promoter. Other strains presented in this Figure were transformed with an empty vector expressing only the LEU2 gene. Transformants were selected on–Leu medium with 150 μM CuSO 4 and replica-plated on–Leu–Ade or–Leu Gal media with 150 μM CuSO 4 . Images were taken after 5 days of incubation of–Ade plates or after 3 passages on Gal plates.

    Journal: PLoS Genetics

    Article Title: Interaction of Prions Causes Heritable Traits in Saccharomyces cerevisiae

    doi: 10.1371/journal.pgen.1006504

    Figure Lengend Snippet: [ SWI + ] prion is a key determinant of nonsense suppression in [ NSI + ] strains. (A) Sedimentation analysis of Swi1(1–297)-YFP protein from the 4-1-1-D931 [ NSI + ] and 1-4-1-1-D931 [ nsi - ] strains expressing pCUP1-SWI1(1–297)-YFP (URA3) plasmid. Soluble (S) and insoluble (I) fractions were obtained as indicated in Materials and Methods. Swi1(1–297)-YFP was detected using monoclonal rabbit primary antibodies against GFP [E385] (ab32146) (Abcam, Great Britain) and ECL Prime Western Blotting Detection Reagent kit (GE Healthcare, USA). Next, SDD-AGE analysis of insoluble fractions of [ NSI + ] and [ nsi - ] strains comprising Swi1(1–297)-YFP was performed. (B) The effects of SWI1 deletion on the [ NSI + ] phenotypic manifestation. SWI1 deletion was obtained as described in Materials and Methods. To express SWI1 , the 11-1-1-D931 [ NSI + ] swi1 Δ strain was transformed with the YGPM19p21 plasmid from the YSC4613 genomic library, containing a genomic fragment encoding SWI1 under the control of its endogenous promoter. Other strains presented in this Figure were transformed with an empty vector expressing only the LEU2 gene. Transformants were selected on–Leu medium with 150 μM CuSO 4 and replica-plated on–Leu–Ade or–Leu Gal media with 150 μM CuSO 4 . Images were taken after 5 days of incubation of–Ade plates or after 3 passages on Gal plates.

    Article Snippet: Proteins fused with CFP, GFP, and YFP were detected using monoclonal rabbit primary antibodies against GFP [E385] (ab32146) (Abcam, Great Britain) and the Amersham ECL Prime Western Blotting Detection Reagent kit (GE Healthcare, USA).

    Techniques: Sedimentation, Expressing, Plasmid Preparation, Western Blot, Transformation Assay, Incubation

    [ SWI + ] and [ PIN + ] prions demonstrate complementary interaction. (A) Comparative analysis of the growth of strains containing combinations of [ prion - ] or [ PRION + ] states for Rnq1 and Swi1 as well as deletions of the corresponding genes. “1”–The [ SWI + ][ pin - ] and [ swi - ][ PIN + ] strains were obtained from the 1-1-D931 [ SWI + ][ PIN + ] strain by deletion with subsequent reintroduction of RNQ1 and SWI1 genes, respectively (see “ Materials and Methods ”). The [ swi - ][ pin - ] strain was obtained from the 1-1-D931 [ SWI + ][ PIN + ] strain by GuHCl curing. “2”–The 26-1-4-1-1-D931 [ swi - ][ PIN + ], 12-1-4-1-1-D931 [ SWI + ][ pin - ], and 16-1-4-1-1-D931 [ SWI + ][ PIN + ] strains were obtained by transformation of the 1-4-1-1-D931 [ swi - ][ pin - ] recipient yeast cells with the 1-1-D931 [ SWI + ][ PIN + ] protein lysates followed by analysis of [ SWI ] and [ PIN ] status of the cells as described in “Materials and Methods”. Images were obtained after 5 days of incubation on–Ade plates with 150 μM CuSO 4 or after 3 passages on Gal plates. (B) Sedimentation analysis of Swi1(1–297)-YFP protein from the 16-1-4-1-1-D931 [ SWI + ][ PIN + ] and 12-1-4-1-1-D931 [ SWI + ][ pin - ] strains expressing the pCUP1-SWI1(1–297)-YFP (URA3) plasmid. Soluble (S) and insoluble (I) fractions were obtained as indicated in Materials and Methods. Swi1(1–297)-YFP was detected using monoclonal rabbit primary antibodies against GFP [E385] (ab32146) (Abcam, Great Britain) and ECL Prime Western Blotting Detection Reagent kit (GE Healthcare, USA). (C) Analysis of the colocalization of Swi1-YFP and Rnq1-CFP aggregates. The cells of the D938 [ SWI + ][ PIN + ] strain were co-transformed with p426GPD–SWI1YFP and pCUP1-RNQ1-CFP(LEU2) plasmids. Transformants were selected on–Ura–Leu selective media with 150 μM CuSO 4 and incubated for 48 h prior to fluorescence microscopy.

    Journal: PLoS Genetics

    Article Title: Interaction of Prions Causes Heritable Traits in Saccharomyces cerevisiae

    doi: 10.1371/journal.pgen.1006504

    Figure Lengend Snippet: [ SWI + ] and [ PIN + ] prions demonstrate complementary interaction. (A) Comparative analysis of the growth of strains containing combinations of [ prion - ] or [ PRION + ] states for Rnq1 and Swi1 as well as deletions of the corresponding genes. “1”–The [ SWI + ][ pin - ] and [ swi - ][ PIN + ] strains were obtained from the 1-1-D931 [ SWI + ][ PIN + ] strain by deletion with subsequent reintroduction of RNQ1 and SWI1 genes, respectively (see “ Materials and Methods ”). The [ swi - ][ pin - ] strain was obtained from the 1-1-D931 [ SWI + ][ PIN + ] strain by GuHCl curing. “2”–The 26-1-4-1-1-D931 [ swi - ][ PIN + ], 12-1-4-1-1-D931 [ SWI + ][ pin - ], and 16-1-4-1-1-D931 [ SWI + ][ PIN + ] strains were obtained by transformation of the 1-4-1-1-D931 [ swi - ][ pin - ] recipient yeast cells with the 1-1-D931 [ SWI + ][ PIN + ] protein lysates followed by analysis of [ SWI ] and [ PIN ] status of the cells as described in “Materials and Methods”. Images were obtained after 5 days of incubation on–Ade plates with 150 μM CuSO 4 or after 3 passages on Gal plates. (B) Sedimentation analysis of Swi1(1–297)-YFP protein from the 16-1-4-1-1-D931 [ SWI + ][ PIN + ] and 12-1-4-1-1-D931 [ SWI + ][ pin - ] strains expressing the pCUP1-SWI1(1–297)-YFP (URA3) plasmid. Soluble (S) and insoluble (I) fractions were obtained as indicated in Materials and Methods. Swi1(1–297)-YFP was detected using monoclonal rabbit primary antibodies against GFP [E385] (ab32146) (Abcam, Great Britain) and ECL Prime Western Blotting Detection Reagent kit (GE Healthcare, USA). (C) Analysis of the colocalization of Swi1-YFP and Rnq1-CFP aggregates. The cells of the D938 [ SWI + ][ PIN + ] strain were co-transformed with p426GPD–SWI1YFP and pCUP1-RNQ1-CFP(LEU2) plasmids. Transformants were selected on–Ura–Leu selective media with 150 μM CuSO 4 and incubated for 48 h prior to fluorescence microscopy.

    Article Snippet: Proteins fused with CFP, GFP, and YFP were detected using monoclonal rabbit primary antibodies against GFP [E385] (ab32146) (Abcam, Great Britain) and the Amersham ECL Prime Western Blotting Detection Reagent kit (GE Healthcare, USA).

    Techniques: Transformation Assay, Incubation, Sedimentation, Expressing, Plasmid Preparation, Western Blot, Fluorescence, Microscopy